CN102586179A - 用于分选带有x或y染色体富集群的精子悬浮液 - Google Patents

用于分选带有x或y染色体富集群的精子悬浮液 Download PDF

Info

Publication number
CN102586179A
CN102586179A CN2012100407786A CN201210040778A CN102586179A CN 102586179 A CN102586179 A CN 102586179A CN 2012100407786 A CN2012100407786 A CN 2012100407786A CN 201210040778 A CN201210040778 A CN 201210040778A CN 102586179 A CN102586179 A CN 102586179A
Authority
CN
China
Prior art keywords
sperm
spermatid
suspension
chromosome
concentration
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN2012100407786A
Other languages
English (en)
Other versions
CN102586179B (zh
Inventor
C·L·路德维格
K·S·克劳利
C·N·格拉维斯
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Monsanto Technology LLC
Inguran LLC
Original Assignee
Monsanto Technology LLC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Monsanto Technology LLC filed Critical Monsanto Technology LLC
Publication of CN102586179A publication Critical patent/CN102586179A/zh
Application granted granted Critical
Publication of CN102586179B publication Critical patent/CN102586179B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/52—Sperm; Prostate; Seminal fluid; Leydig cells of testes
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0205—Chemical aspects
    • A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0205—Chemical aspects
    • A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
    • A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0608—Germ cells
    • C12N5/061—Sperm cells, spermatogonia
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0608—Germ cells
    • C12N5/0611—Primordial germ cells, e.g. embryonic germ cells [EG]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis

Abstract

本发明公开了包含运动抑制剂的精子细胞悬浮液。此类悬浮液中含有的细胞更能耐受与将精子细胞分选为性别富集群过程一般相关的多个步骤，从而使分选后的组合物中活精子或运动性的精子数目增多。本发明也公开了形成此类细胞悬浮液的方法以及对精子细胞染色的方法。

Description

用于分选带有X或Y染色体富集群的精子悬浮液
本申请为2005年3月29日提交的、发明名称为“用于分选带有X或Y染色体富集群的精子悬浮液”的PCT申请PCT/US2005/010481的分案申请，所述PCT申请进入中国国家阶段的日期为2006年11月28日，申请号为200580017370.5。
发明领域
本发明主要涉及分选精子细胞的方法。更具体而言，本发明涉及制备精子细胞运动性相对于体内射精精子降低的悬浮液，更具体而言是运动性暂时降低；所述悬浮液可用于如将精子细胞分选为带有X或Y染色体的富集精子细胞群的方法。
发明背景
通过人工授精(AI)使动物受精以及体外受精后的胚胎移植已经是一项成熟的技术。在家畜产业中，能够对繁殖结果施加影响而使其后代具有一种或多种所需特征，有明显的优势。例如，乳制品产业中如能预先选择后代有利于雌性而保证奶牛的产量，则会带来经济效益。将精子分离为富集的带有X和Y染色体的细胞群，即所谓的性别富集精液或性别富集精子，便是达到后代预选目的的方法之一。
为得到性别富集精液，必须将精子细胞以染料染色，然后分选为带有X和Y染色体的细胞。染色和分选过程的每一步对于精子细胞而言都是应激，会导致精子细胞的存活力或运动性，特别是前向性运动性降低。
Salisbury等人描述了将公牛射出的精液直接收集至稀释液中的技术，所述稀释液可抑制细胞的运动性并防止其从周围精浆中吸收糖。当收集射出精液至稀释液中，且通过充气100％的CO2替换液体上方的气相时，射出精液内的细胞会丧失运动性。只要将细胞留存于该稀释液中且排除空气，就可以保证其在室温下数小时不运动，而在5℃时可维持至少8天。
发明概述
本发明的多方面之一涉及精子悬浮液，所述精子悬浮液可用于如将精子分选为带X或Y染色体的富集精子群的方法。
因此简言之，本发明涉及含有活精子以及下调精子糖摄取的组合物的精子细胞悬浮液，所述悬浮液中的精子浓度小于约1×106精子/ml或至少为1×108精子/ml。
本发明还涉及含有不运动活精子的精子细胞悬浮液，所述悬浮液中的精子浓度小于约1×106精子/ml或至少为1×108精子/ml。
本发明还涉及含有活精子的精子细胞悬浮液，所述精子相对于同一种系体内射出的精子更具附睾中精子的运动性特点，所述悬浮液中的精子浓度小于约1×106精子/ml或至少为1×108精子/ml。
本发明还涉及含有活精子、钾、并任选地含有钠的精子细胞悬浮液，所述悬浮液中的精子浓度至少为1×108精子/ml，而钾与钠的摩尔比大于各1∶1。
本发明还涉及含有活精子、下调精子糖摄取的组合物、以及DNA选择性染料的精子细胞悬浮液。
本发明还涉及含有不运动活精子和DNA选择性染料的精子细胞悬浮液。
本发明还涉及含有活精子和DNA选择性染料的精子细胞悬浮液，所述精子相对于同一种系体内射出的精子，其代谢率和运动性更具附睾中精子的特点。
本发明还涉及含有不运动活精子的精子细胞悬浮液，所述精子的DNA上结合有DNA选择性染料。
本发明还涉及含有活精子的精子细胞悬浮液，相对于同一种系体内射出的精子而言，所述精子代谢率和运动性更具附睾中精子的特点，且其DNA上还结合有DNA选择性染料。
本发明还涉及对精子细胞染色的方法，所述方法包括形成这样的染色混合物，其含有完整的活精子细胞、运动抑制性量的钾，以及DNA选择性染料。
本发明还涉及形成用于流式细胞术方法的精子细胞悬浮液的方法，所述方法包括将精子细胞源与抑制精子细胞运动性的组合物混合，以形成精子细胞悬浮液，所述悬浮液中精子细胞浓度小于约1×106精子细胞/ml或至少为1×108精子细胞/ml。
本发明还涉及形成用于流式细胞术方法的精子细胞悬浮液的方法，所述方法包括将哺乳动物射出精液收集至含有抑制性量的运动抑制剂的缓冲液中以形成精子悬浮液，所述悬浮液中含有精子浓度小于约1×106精子细胞/ml或至少为1×108精子细胞/ml。
本发明的其它方面和特征是显而易见的，部分将于后文说明。
附图简述
图1以图表显示了实施例1中进行研究的结果，其中测量了于28℃含10mM丙酮酸盐的TCA或二氧化碳气层包裹的含10mM丙酮酸盐的TCA中，用600μM Hoechst 33342染料染色的精子细胞的前向性运动百分比。
图2以图表显示了实施例1中进行研究的结果，其中测量了于28℃含有10mM丙酮酸盐的TCA或pH 7.3基于碳酸盐的抑制剂中，用600μMHoechst 33342染料染色的精子细胞的前向性运动百分比。
图3以图表显示了实施例1中进行研究的结果，其中测量了于28℃含有10mM丙酮酸盐的TCA或pH 6.2基于碳酸盐的抑制剂中，用600μMHoechst 33342染料染色的精子细胞的前向性运动百分比。
图4以图表显示了实施例2中进行研究的结果，其中测量了于28℃含10mM丙酮酸盐的TCA且随后用含有10mM丙酮酸盐的TCA或pH 6.2基于碳酸盐的抑制剂1比3稀释的情况下，用1000μM Hoechst 33342染料染色的精子细胞的前向性运动百分比。
图5以图表显示了实施例2中进行研究的结果，其中测量了于28℃(1)含有10mM丙酮酸盐的TCA且用相同溶液1比3稀释的情况下或(2)pH 7.3基于碳酸盐的缓冲液且用pH 6.2基于碳酸盐的抑制剂1比3稀释的情况下，用1000μM Hoechst 33342染料染色的精子细胞的前向性运动百分比。
图6以图表显示了实施例2中进行研究的结果，其中测量了于28℃含有10mM丙酮酸盐的TCA或pH 6.2基于碳酸盐的抑制剂中，用1000μMHoechst 33342染料染色的精子细胞的前向性运动百分比。
图7以图表显示了实施例3中进行研究的结果，其中测量了于41℃含有10mM丙酮酸盐的TCA且随后用含有10mM丙酮酸盐的TCA或pH6.2基于碳酸盐的抑制剂1比3稀释的情况下，用300μM Hoechst 33342染料染色的精子细胞的前向性运动百分比。
图8以图表显示了实施例3中进行研究的结果，其中测量了于41℃(1)含有10mM丙酮酸盐的TCA且用相同溶液1比3稀释的情况下或(2)pH 7.3基于碳酸盐的缓冲液且用pH 6.2基于碳酸盐的抑制剂1比3稀释的情况下，用300μM Hoechst 33342染料染色的精子细胞的前向性运动百分比。
图9以图表显示了实施例3中进行研究的结果，其中测量了于41℃含有10mM丙酮酸盐的TCA或pH 6.2基于碳酸盐的抑制剂中，用300μMHoechst 33342染料染色的精子细胞的前向性运动百分比。
图10以图表显示了实施例4中进行研究的结果，其中测量了于41℃TCA缓冲液或含有10mM丙酮酸盐的TCA缓冲液中，用400μM Hoechst33342染料染色的精子的前向性运动百分比。
图11以图表显示了实施例5中进行研究的结果，其中测量了于41℃TCA缓冲液或含有10μM维生素K的TCA缓冲液中，用400μM Hoechst33342染料染色的精子的前向性运动百分比。
图12以图表显示了实施例6中进行研究的结果，其中测量了于41℃TCA缓冲液或含有100μM维生素K的TCA缓冲液中，用400μM Hoechst33342染料染色的精子的前向性运动百分比。
图13以图表显示了实施例7中进行研究的结果，其中测量了于41℃TCA缓冲液或含1mM硫辛酸的TCA缓冲液中，用400μM Hoechst 33342染料染色的精子的前向性运动百分比。
图14以图表显示了实施例8中进行研究的结果，其中测量了于28℃TCA缓冲液或含有10mM丙酮酸盐的TCA缓冲液中，用600μM Hoechst33342染料染色的精子的前向性运动百分比。
图15以图表显示了实施例9中进行研究的结果，其中测量了于28℃TCA缓冲液或含有100μM维生素K的TCA缓冲液中，用600μM Hoechst33342染料染色的精子的前向性运动百分比。
图16以图表显示了实施例10中进行研究的结果，其中测量了于28℃TCA缓冲液或含1mM硫辛酸的TCA缓冲液中，用600μM Hoechst 33342染料染色的精子的前向性运动百分比。
图17以图表显示了实施例11中进行研究的结果，其中测量了于28℃TCA缓冲液、含有2.5mM丙酮酸盐的TCA缓冲液、含有10mM丙酮酸盐的TCA缓冲液、含有25mM丙酮酸盐的TCA缓冲液以及含有50mM丙酮酸盐的TCA缓冲液中，用600μM Hoechst 33342染料染色的精子的前向性运动百分比。
图18以图表显示了实施例12中进行研究的结果，其中测量了于28℃TCA缓冲液或含有10mM丙酮酸盐的TCA缓冲液中，用20μM SYBR-14染料染色的精子的前向性运动百分比。
图19以图表显示了实施例13中进行研究的结果，其中测量了于28℃TCA缓冲液或含有10mM丙酮酸盐的TCA缓冲液中，用100μM BBC染料染色的精子的前向性运动百分比。
图20以图表显示了实施例14中进行研究的结果，其中测量了于28℃TCA缓冲液或含有10mM丙酮酸盐的TCA缓冲液中，用200μM BBC染料染色的精子的前向性运动百分比。
发明详述
令人惊讶的是，已经确定相对于体内射精精子(相同种系)运动性降低的精子，往往更能耐受多个与分选精子细胞为带有X或Y染色体的精子富集群通常相关的步骤。因此在优选实施方案中，可制备用于人工授精的性别富集精子群，所述精子群在染色后或分选后的组合物中活细胞数目增多或运动精子(特别是前向运动性精子)数目增多。
根据本发明的方法，形成含有精子和一种或多种抑制精子运动性的组合物的悬浮液，有时也称其为分散体系；此种运动性受抑的状态有时被称之为不运动或精子静止。一般而言，悬浮液中含有精子的密度为约1×103精子/ml至约5×1010精子/ml悬浮液。例如，在一个实施方案中，悬浮液中可含有“相对低”密度的精子，即精子密度少于约1×107精子/ml，优选少于约1×106精子/ml，更优选约1×103至约5×106精子/ml，更优选约1×103至约1×106精子/ml，甚至更优选1×104至约1×105精子/ml，最优选约1×105精子/ml悬浮液。在可选的实施方案中，悬浮液中可含有“中间”密度的精子，即密度为约1×107至约1×108精子/ml悬浮液。而在另一个实施方案中，悬浮液中可含有“相对高”密度的精子，即精子密度至少为约1×108精子/ml，优选约1×108至约5×1010精子/ml，更优选约1.5×108至约2×1010精子/ml，甚至更优选1.5×108至约2×108精子/ml，更加优选约1.5×108精子/ml悬浮液。因此例如，在一个实施方案中，悬浮液中可含有至少约1.25×108、至少约1.5×108、至少约1.75×108、至少约2×108、至少约2.25×108、至少约2.5×108、至少约2.75×108，或甚至至少约3×108精子/ml悬浮液。在可选的实施方案中，悬浮液中可含有少于约9×105、少于约7×105、少于约5×105、少于约2×105、少于约1×105、少于约1×104，或甚至少于约1×103精子/ml悬浮液。
精子悬浮液中的精子密度取决于若干考虑，包括随后将用于富集或分选精子细胞的方法。例如，可以利用如下文详述的流式细胞术分选精子细胞。在这样的情况下，缓冲精子悬浮液一般含“中间”或“相对高”密度的精子。其它分选或富集技术可能在精子密度较低时更利于使用，例如标有标记(如此处所述的染料和标记)的“相对低”密度的精子。
在优选实施方案中，本发明悬浮液中的精子在某些方面表现出附睾中精子的特征性行为；例如，精子的不运动性，和/或相对于洗涤或刚射出的精子而言，其内呼吸速率更低而有氧糖酵解的速率更高。最好受抑精子在与抑制剂分离后，其运动性能够表现出射出精子的特征性行为(而不具有附睾精子的特点)；而在一个实施方案中，其运动性和呼吸都可具有如射出精子的特征性表现。
例如在一个实施方案中，通过HTM-IVOS精子分析(Hamilton-ThorneHTM-IVOS计算机辅助精子分析系统，Hamilton-Thorne Research，Beverly MA)测量，运动抑制剂可使分散体系中精子细胞的路径速度、前向性速度或两者同时相对于相同种系刚射出精液中精子细胞的路径速度、前向性速度或两者同时，降低至少约50％。优选通过HTM-IVOS精子分析测量，运动抑制剂可使分散体系中精子细胞的路径速度、前向性速度或两者同时相对于相同种系刚射出精液中精子细胞的路径速度、前向性速度或两者同时，降低至少约60％。更优选通过HTM-IVOS精子分析测量，运动抑制剂可使分散体系中精子细胞的路径速度、前向性速度或两者同时相对于相同种系刚射出精液中精子细胞的路径速度、前向性速度或两者同时，降低至少约70％。还更优选通过HTM-IVOS精子分析测量，运动抑制剂可使分散体系中精子细胞的路径速度、前向性速度或两者同时相对于相同种系刚射出精液中精子细胞的路径速度、前向性速度或两者同时，降低至少约80％。甚至更优选通过HTM-IVOS精子分析测量，运动抑制剂可使分散体系中精子细胞的路径速度、前向性速度或两者同时相对于相同种系刚射出精液中精子细胞的路径速度、前向性速度或两者同时，降低至少约90％。甚至更优选通过HTM-IVOS精子分析测量，运动抑制剂可使分散体系中精子细胞的路径速度、前向性速度或两者同时相对于相同种系刚射出精液中精子细胞的路径速度、前向性速度或两者同时，降低至少约95％。最优选通过HTM-IVOS精子分析测量，运动抑制剂可使分散体系中精子细胞的路径速度、前向性速度或两者同时相对于相同种系刚射出精液中精子细胞的路径速度、前向性速度或两者同时，降低至少约99％。
除抑制性缓冲液外或可替代之的是，也可单独降低精子细胞的温度或紧邻精子细胞的环境温度(即精子分散体系)以影响细胞的运动性。此种降温一般会增加不运动性。此外，例如，精子细胞或精子分散体系温度的降低会导致用于诱导不运动性的抑制剂的浓度降低。因此，精子分散体系可处于不超过5℃的温度；优选在约0℃至约5℃之间；更优选为约3℃至约5℃之间；而最优选为约5℃。或者精子分散体系的温度可介于约4℃至约50℃范围内；优选为约7℃至约43℃；更优选为约10℃至约39℃；还更优选为约15℃至约30℃；甚至更优选为约17℃至约25℃；而最优选为约18℃。然而，优选不将精子细胞暴露于会对细胞存活力造成实质不良影响的温度下。
抑制剂可以为一系列对精子运动性具有抑制作用的组合物中的任意一种。此类组合物包括如钠/钾ATP酶抑制剂，如乌本苷；含有钾离子的组合物；以及含有钾离子和钠离子的组合物。例如，悬浮液中相对高浓度的钾离子会抑制精子运动性。因此一般而言，优选悬浮液中含有钾离子源且悬浮液中的钾浓度至少为约0.05mol/L。更优选钾浓度为至少约0.05mol/L至约0.5mol/L。还更优选钾浓度为至少约0.1mol/L至约0.3mol/L。最优选钾浓度为约0.173摩尔/升。此类悬浮液一般(但非必须)也含有钠离子源。当含有钠离子时，钾和钠的摩尔比一般等于或大于各1∶1。优选钾和钠的摩尔比为至少约1.25∶1。更优选钾和钠的摩尔比为至少约1.5∶1。还更优选钾和钠的摩尔比为至少约1.75∶1。还更优选钾和钠的摩尔比为至少约1.78∶1。在一个具体的实施方案中，钾和钠的摩尔比为至少约2∶1。而在另一个实施方案中，钾和钠的摩尔比为至少约3∶1。又在另一个实施方案中，钾和钠的摩尔比为至少约4∶1。又在另一个实施方案中，钾和钠的摩尔比为至少约5∶1。又在另一个实施方案中，钾和钠的摩尔比为至少约6∶1。又在另一个实施方案中，钾和钠的摩尔比为至少约7∶1。又在另一个实施方案中，钾和钠的摩尔比为至少约8∶1。
精子悬浮液中还可额外含有能够下调糖摄取的离子或二氧化碳源。在该实施方案中，二氧化碳的来源可以为如一种或多种碳酸盐。在目前优选的一个实施方案中，精子悬浮液含有NaHCO3和KHCO3，由此提供钾和钠离子源以及二氧化碳分压。例如，在目前优选的一个实施方案中，精子悬浮液含有NaHCO3和KHCO3的水性溶液，优选NaHCO3、KHCO3和C6H8O7·H2O的水溶液；一般而言，分散体系中KHCO3的浓度可为至少约0.05mol/L。更优选KHCO3的浓度为至少约0.05mol/L至约0.5mol/L。还更优选KHCO3的浓度为至少约0.1mol/L至约0.3mol/L。在特别优选的实施方案中，如Salisbury & Graves，J.Reprod.Fertil.，6：351-359(1963)中公开的那样，利用含0.097mol/L NaHCO3、0.173mol/L KHCO3、0.090mol/L C6H8O7·H2O水溶液的抑制性缓冲液形成悬浮液。只要精子细胞暴露于运动抑制剂，它们一般都会保持静止。
当分散体系中含有C6H8O7·H2O时，KHCO3与NaHCO3间的摩尔比可如上所述。KHCO3与C6H8O7·H2O间的摩尔比一般等于或大于各1∶1，但一般不会超过摩尔比8∶1。优选KHCO3与C6H8O7·H2O间的摩尔比为至少约1.25∶1。还更优选KHCO3与C6H8O7·H2O间的摩尔比为至少约1.5∶1。还更优选KHCO3与C6H8O7·H2O间的摩尔比为至少约1.75∶1。在一个具体实施方案中，KHCO3与C6H8O7·H2O间的摩尔比为至少约1.78∶1。在另一具体实施方案中，KHCO3与C6H8O7·H2O间的摩尔比为至少约2∶1。而在另一实施方案中，KHCO3与C6H8O7·H2O间的摩尔比为至少约3∶1。又在另一实施方案中，KHCO3与C6H8O7·H2O间的摩尔比为至少约4∶1。又在另一实施方案中，KHCO3与C6H8O7·H2O间的摩尔比为至少约5∶1。又在另一实施方案中，KHCO3与C6H8O7·H2O间的摩尔比为至少约6∶1。又在另一实施方案中，KHCO3与C6H8O7·H2O间的摩尔比为至少约7∶1。又在另一实施方案中，KHCO3与C6H8O7·H2O间的摩尔比为至少约8∶1。在特别优选的实施方案中，如Salisbury & Graves，J.Reprod.Fertil.，6：351-359(1963)中所公开的那样，利用含0.097mol/L NaHCO3、0.173mol/L KHCO3、0.090mol/L C6H8O7·H2O的水溶液的抑制性缓冲液形成悬浮液。只要精子细胞暴露于运动抑制剂，它们一般都会保持静止。
迄今为止的实验证据还提示，如果将精子细胞悬浮液保存在相对于空气二氧化碳分压增高的大气下，可以改善精子细胞的整体健康和其它生命特征。在优选实施方案中，悬浮液上方的大气中二氧化碳分压值为至少0.9；更优选为至少约0.95。
可以将细胞与运动性抑制剂分离并将其暴露于空气中，使静止细胞还原为活性状态。此外，还可以通过在生理盐水中(salisbury等人，1963)或诸如TCA缓冲液或PBS的缓冲液中稀释细胞来起始活性状态。一般而言，根据HTM-IVOS精子分析测量，至少约20％、优选至少约50％、更优选至少约60％、还更优选至少约70％、甚至更优选至少约80％、甚至更优选至少约90％、还更优选至少约95％、而最优选至少约99％回复活性状态的细胞(即再活化细胞)具有这样的路径速度、前向性速度或两者兼具：即分别为与运动性抑制剂混合前的精子细胞(即刚射出的精子细胞)的路径速度、前向性速度或两者的至少约50％，优选至少约60％，更优选至少约70％，还更优选至少约80％，甚至更优选至少约90％，甚至更优选至少约95％，而最优选至少约99％。
一般而言，细胞分选方法包括一系列分离步骤，即细胞样本的收集、细胞染色、细胞分选、分选细胞的收集，并任选地包括分选细胞的冷冻伸展(cryoextension)。形成的精子悬浮液中最好包含运动性抑制剂，或在一个或多个这些步骤中采用之。
细胞样品的收集
可以收集牛、猪、马或其它哺乳动物完整的活精子细胞并将之与运动性抑制剂接触。已知众多收集活精子的方法，例如，包括手握法、使用人工阴道以及电刺激射精。例如，可以将一般每毫升含有5亿至
Figure BDA0000137094600000101
细胞的牛精液样品，直接由来源哺乳动物收集于含有运动性抑制剂的容器中，以形成精子悬浮液。可选地，也可以将精子样品收集于空容器中，随后在收集的数分钟至数小时内将之与运动性抑制剂接触以形成精子悬浮液。
除缓冲物外，精子悬浮液中也可含有一系列添加剂以增强精子存活力。例示性的添加剂包括蛋白质源、抗生素以及调节细胞内和/或细胞外氧化/还原反应的组合物。
例示性的蛋白质源包括卵黄、卵黄提取物、乳(包括热匀浆和脱脂乳)、乳提取物、大豆蛋白、大豆蛋白提取物、血清白蛋白、牛血清白蛋白、人血清替代物增补剂以及它们的组合。白蛋白特别是牛血清白蛋白(BSA)，是优选的蛋白质源。例如，若含有BSA，则精子悬浮液中的BSA含量可少于约5.0％(w/v)，优选少于约2％(w/v)，更优选少于约1％(w/v)，而最优选其量为约0.1％(w/v)。
单独使用蛋白质源(如BSA)，可能会起始悬浮液中百分之一精子细胞的获能过程。该过程优选发生于雌性生殖道内。因此，为了抑制稀释过程以及随后的染色和分选期间获能的起始，精子悬浮液中可含有备选的蛋白质源或蛋白质替代物。所述备选蛋白质源或蛋白质替代物除了能够抑制精子悬浮液中更大百分比的细胞起始获能，还具有典型蛋白质源(如BSA)的优点。备选白质源的实例包括人血清替代物增补剂(SSS)(IrvineScientific，Santa Ana，CA)以及胆固醇强化的BSA，而蛋白质替代物的实例包括聚乙烯醇，例如一般分子量为约30,000至约60,000的低至中粘度聚乙烯醇。一般而言，若含有这些组合物，其含量与上述BSA的量相同；缓冲物或缓冲溶液中总白蛋白含量一般不超过约5.0％(w/v)。
例如，调节细胞内和/或细胞外氧化/还原反应的例示性组合物包括丙酮酸盐、维生素K、硫辛酸、谷胱甘肽、黄素、醌类、超氧化物歧化酶(SOD)以及SOD模拟物。若精子悬浮液中含有此类组合物，则其浓度足以产生保护性作用而不会对精子健康产生不良影响。例示性的浓度范围包括从约10μM至约20mM，取决于诸如所用的具体组合物或悬浮液中的精子浓度等因素。例如，精子悬浮液中丙酮酸盐的浓度可为约1mM至约20mM，优选为约5mM至约15mM，更优选为约10mM。精子悬浮液中维生素K的浓度可为约1μM至约100μM，优选为约10μM至约100μM，更优选为约100μM。精子悬浮液中硫辛酸的浓度可为约0.1mM至约1mM，优选为约0.5mM至约1mM，而更优选为约1mM。
精子悬浮液中可含有抗生素以抑制细菌的生长。例示性的抗生素包括例如泰乐菌素、庆大霉素、林可霉素、壮观霉素、利高霉素Linco-
Figure BDA0000137094600000111
(林可霉素羟氯化物-壮观霉素)、青霉素、链霉素、替卡西林(ticarcillin)或其任意组合。若含有抗生素，则其浓度可为每ml精液中约50μg至约800μg(不论精液是纯净的、缓冲性的或含有其它物质，例如此处提及的任一种添加剂)。Certified Semen Services(CSS)和美国国家动物育种协会(NAAB)已经颁布了有关抗生素在精子收集和使用中应用的纲要。
可以向精子分散体系中加入生长因子，以协助维持精子细胞的存活力。例示性的生长因子包括如转化生长因子(“TGF”)(例如TGF β-1和TGFβ-2)，以及胰岛素样生长因子(“IGF”)(例如IGF-1)。一般而言，精子分散体系中的TGF可以TGF β-1的形式出现，其浓度为约0.1ng/L至约10μg/L，或为TGF β-2的形式，其浓度为约0.1ng/L至约200ng/L；而IGF在精子分散体系中可以为IGF-1形式，其浓度为约0.1ng/L至约50μg/L。此类生长因子的用途为本领域所公知，并公开于如美国专利申请公开号2003/0157473中，其内容在此处引用作为参考。
一旦收集到细胞，可将其于室温下静止状态保存数小时、低温下(如5℃)保存数周，或在下文所讨论的冰冻伸展剂中保存数月。优选细胞上方的大气如上所述含有较高的二氧化碳分压。或者，收集的细胞还可在数小时内用于如育种方法、染色方法或分选方法。
细胞染色
运动性抑制剂可用于使细胞在细胞染色期间不运动。一般而言，精子细胞染色方法中可包括形成染色混合物(有时称之为标记混合物)，其中含有完整的活精子细胞、运动性抑制剂和染料(有时称之为标记)。在本发明的这一方面，可以将运动性抑制剂与精子细胞接触以形成精子悬浮液，然后将精子悬浮液与DNA选择性染料相接触。在该实施方案中，精子来源可为纯精子，或者为通过离心或使用其它手段分离精子级分而得到的含精子的精液衍生物。
在可选的实施方案中，染料可与运动性抑制剂相混合，从而形成染料溶液。因此，例如，纯固体(包括可自由流动的粉末)或液体组合物形式的染料可与抑制剂组合形成染料溶液，所述溶液可随后与纯精子、精子悬浮液或含精子的精液衍生物混合。
任何情况下，只要精子细胞保存在抑制剂中，一般都会保持静止(Salisbury等人，1963)。然而，优选将染色混合物保存在二氧化碳分压相对于空气而言增高的大气中；例如，一般优选染色混合物上方的大气中含99％+的CO2。
可将染色混合物的pH保持在一定pH值范围内的任意一点；一般而言，该范围为约5.0至约9.0。例如，可使染色混合物保持在“微酸性”pH下，即约5.0至约7.0。在该实施方案中，优选pH为约6.0至约7.0，更优选为约6.0至约6.5，而最优选为约6.2。或者，还可使染色混合物保持在“微碱性”pH下，即约7.0至约9.0。在该实施方案中，优选pH为约7.0至约8.0，更优选为约7.0至约7.5，而最优选为约7.3。
如以前在美国专利号5,135,759和WO 02/41906中所述，可使用一种或多种可经UV或可见光激发的DNA选择性染料形成染色混合物。经紫外光(UV)激发的选择性染料的实例包括Hoechst 33342和Hoechst 33258，二者均可由Sigma-Aldrich(St.Louis，MO)购得。经可见光激发的例示性选择性染料包括可购于Molecμlar Probe，Inc.(Eugene，OR)的SYBR-14，以及WO02/41906中所描述的双苯甲亚胺-
Figure BDA0000137094600000131
缀合物6-{[3-((2Z)-2-{[1-(二氟硼烷基)-3，5-二甲基-1H-吡咯-2-基]亚甲基}-2H-吡咯-5-基)丙酰基]氨基}-N-[3-(甲基{3-[({4-[6-(4-甲基哌嗪-1-基)-1H，3’H-2，5’-双苯并咪唑-2’-基]苯氧基}乙酰基)氨基]丙基}氨基)丙基]己酰胺(“BBC”)。这些染料中的每一种都可以单独使用或者联合使用；或者，也可以将其它具有细胞渗透性的UV和可见光激发的染料单独或与前述染料联合使用，只要所述染料在可进行别处所述分选的浓度下不会对精子细胞的存活力产生不可接受的不良影响。
或者，还可利用荧光聚酰胺形成染色混合物，更具体而言是利用缀合有荧光标记或报告子的聚酰胺。此类标记在结合于核酸时会发出荧光。连接有荧光标记或报告子的聚酰胺实例包括，如Best等人，Proc.Natl.Acad.Sci.USA，100(21)：12063-12068(2003)；Gygi等人，Nucleic Acids Res.，30(13)：2790-2799(2002)；美国专利号5,998,140；美国专利号6,143,901以及美国专利号6,090,947中所公开的那些，其中每一篇内容均在此处引用作为参考。
荧光核苷酸序列也可用于标记精子细胞。此类核苷酸序列在与含有靶序列或互补序列的核酸杂交时会发出荧光，但在非杂交状态时则不会发出荧光。此类核苷酸公开见于如美国专利申请公开号2003/0113765(此处引用作为参考)。
性别特异性抗体也可用于标记染色混合物中的精子细胞。例如，在该实施方案中，性别特异性抗体可缀合于荧光部分(或等价报告分子)。由于抗体结合的抗原只存在于带有X染色体或Y染色体的细胞上，根据其荧光(相对于无荧光的未标记细胞)能够选择性地鉴定此类细胞。此外，可以同时使用分别与不同荧光部分相连的一种以上的性别特异性抗体。从而能根据带有X染色体或Y染色体的细胞间的不同荧光，对之进行区分。
也可使用发光的色彩选择性纳米晶体标记染色混合物中的精子细胞。这些颗粒也称为量子点，正如美国专利号6,322,901和美国专利号6,576,291所述为本领域所公知(均在此处引用作为参考)。这些纳米晶体已被缀合于多种生物学材料，包括如肽、抗体、核酸、链霉亲和素及多糖(见如美国专利号6,207,392、6,423,551、5,990,479和6,326,144，均在此处引用作为参考)，且已用于检测生物学靶标(见如美国专利号6,207,392和6,247,323，两者均在此处引用作为参考)。
染色混合物中DNA选择性染料或任何其它类型染料的浓度优选为一系列变量的函数，所述变量包括细胞对于所选染料的渗透性、染色混合物的温度、完成染色所需要的时间量、以及在随后的分选步骤中所期望的富集程度。一般而言，优选的染料浓度足以在适度短的时间内达到所期望的染色程度，而不会对精子存活力产生实质的不良影响。例如，Hoechst33342、Hoechst 33258、SYBR-14或BBC在染色混合物中的浓度一般在约0.1μM至约1.0M之间，优选为约0.1μM至约700μM，更优选为约100μM至约200μM。在特别优选的实施方案中，Hoechst 33342、Hoechst33258、SYBR-14或BBC在染色混合物中的浓度一般在约400μM至约500μM，最优选为约450μM。因此，在一种染色条件设置下，Hoechst 33342的浓度优选为约100μM。在另一种染色条件设置下Hoechst 33342的浓度为约150μM。又在另一种染色条件设置下，浓度优选为约200μM。而在另一种染色条件设置下，Hoechst 33342的浓度最优选为约450μM。
又例如，荧光聚酰胺的浓度(例如美国申请公开号2001/0002314中所述的那些)，一般为约0.1μM至约1mM，优选约1μM至约1mM，更优选约5μM至约100μM，甚至更优选约10μM。
染色混合物中也可任选地包含可增强精子存活力的添加剂。例示性的添加剂包括如上文“细胞样品的收集”中已经讨论过的抗生素、生长因子或调节细胞内和/或细胞外氧化/还原反应的组合物。可据此向收集液中添加此类添加剂。
染色混合物一旦形成，可以在一系列温度范围中的任意温度下保存；一般而言，温度范围为约4℃至约50℃。例如，可以将染色混合物保存于“相对低”的温度下，即温度为约4℃至约30℃；在该实施方案中，温度优选为约20℃至约30℃，更优选为约25℃至约30℃，最优选为约28℃。或者也可将染色混合物保存于“中间”温度范围内，即温度为约30℃至约39℃；在该实施方案中，温度优选为约34℃至约39℃，更优选为约37℃。此外，还可将染色混合物保存于“相对高”的温度范围内，即温度为约40℃至约50℃；在该实施方案中，温度优选为约40℃至约45℃，更优选为约40℃至约43℃，最优选为约41℃。对于优选温度的选择一般取决于一系列变量，包括如细胞对所用染料的渗透性、染色混合物中染料的浓度、细胞在染色混合物中留存的时间、以及在分选步骤中所期望的富集程度。
精子细胞摄取染色混合物中的染料可以持续足够长的一段时间，以达到所期望程度的DNA染色。所述时间一般足以使染料结合于精子细胞的DNA，从而能根据带有X和Y染色体的精子细胞间不同的且可测量的荧光强度，将两者分选开来。一般而言，这一时间不会超过约160分钟，优选不超过约90分钟，还更优选不超过约60分钟，而最优选为约5分钟至约40分钟。
因此，在一个实施方案中，形成含有精子细胞、运动性抑制剂和浓度为约100μM至约200μM的染料的染色混合物，且将该染色混合物于约41℃的温度保持一段时间。在另一个实施方案中，运动性抑制剂于每25ml纯水中含有0.204g NaHCO3、0.433g KHCO3和0.473g C6H8O7·H2O(NaHCO3 0.097mol/L、KHCO3 0.173mol/L、C6H8O7·H2O 0.090mol/L的水溶液)。
在另一个实施方案中，形成含有精子细胞、运动性抑制剂和浓度为约400μM至约500μM的染料的染色混合物，且将该染色混合物于约41℃的温度保持一段时间。在另一个实施方案中，染料浓度为450μM。在另一个实施方案中，运动性抑制剂于每25ml纯水中含有0.204g NaHCO3、0.433g KHCO3和0.473g C6H8O7·H2O(NaHCO3 0.097mol/L、KHCO30.173mol/L、C6H8O7·H2O 0.090mol/L的水溶液)。
又在另一个实施方案中，形成含有精子细胞、运动性抑制剂和浓度为约100μM至约200μM的染料的染色混合物，且将该染色混合物于约28℃的温度保持一段时间。在另一个实施方案中，运动性抑制剂于每25ml纯水中含有0.204g NaHCO3、0.433g KHCO3和0.473g C6H8O7·H2O(NaHCO3 0.097mol/L、KHCO3 0.173mol/L、C6H8O7·H2O 0.090mol/L的水溶液)。
而在另一个实施方案中，形成含有精子细胞、运动性抑制剂和浓度为约400μM至约500μM的染料的染色混合物，且将该染色混合物于约28℃的温度下保持一段时间。在另一个实施方案中，染料浓度为450μM。在另一个实施方案中，运动性抑制剂于每25ml纯水中含有0.204g NaHCO3、0.433g KHCO3和0.473g C6H8O7·H2O(NaHCO3 0.097mol/L、KHCO30.173mol/L、C6H8O7·H2O 0.090mol/L的水溶液)。
分选
运动性抑制剂也可用于在分选精子细胞期间使细胞不运动。一般而言，一旦根据本发明将精子染色，可以根据任何已知的可依据荧光分离的方法分选之。常用且公知的方法包括流式细胞术体系，如美国专利号5,135,759、5,985,216、6,071,689、6,149,867及6,263,745；国际专利公开号WO99/33956和WO01/37655；美国专利申请系列号10/812,351及其相应的国际专利公开号WO 2004/088283中所例示和描述的那些，此处引用其内容作为参考)。当根据这些方法分选时，将精子作为样品液引入流式细胞仪的管口中。因此在一个实施方案中，样品液可包含有染色精子细胞和运动性抑制剂。
与之类似，用于在穿行细胞仪时包裹样品液流的鞘液中也可含有运动性抑制剂。一般而言，利用加压空气或注射泵将鞘液引入流式细胞仪的管口。优选的加压空气为二氧化碳或氮气，更优选氮气。或者，加压空气也可为二氧化碳，但在这种情况下，必须注意尽量减少气泡。
样品液或鞘液中也可任选地含有添加剂，如上文“细胞样品的收集”中讨论过的抗生素、调节细胞内和/或细胞外氧化/还原反应的组合物或生长因子。可据此将这些添加剂中的每一种加入任一种液体中。
分选细胞的收集
一旦完成分选，将分选细胞收集于含有收集液的容器内。一般而言，(采用)收集液的目的包括缓冲收集容器对精子细胞的影响，或为细胞提供液体支持。
在一个实施方案中，收集液中可含有运动性抑制剂和蛋白质源。若含有蛋白质源，则其可为任何不会干扰精子细胞存活力并与所用的运动性抑制剂相容的蛋白质源。常用蛋白质源的实例包括乳(包括热匀浆和脱脂乳)、乳提取物、卵黄、卵黄提取物、大豆蛋白和大豆蛋白提取物。此类蛋白质的使用浓度可为约1％(v/v)至约30％(v/v)，优选约10％(v/v)至约20％(v/v)，更优选约10％(v/v)。
收集液中也可任选地含有添加剂，如上文“细胞样品的收集”中讨论过的抗生素、生长因子或调节细胞内和/或细胞外氧化/还原反应的组合物。可据此将这些添加剂中的每一种加入收集液中。
因此，在某一实施方案中，收集液为含有0.097mol/L NaHCO3、0.173mol/L KHCO3、0.090mol/L C6H8O7·H2O以及10％(v/v)卵黄的水溶液，pH为约6.2，更优选pH为约7.0，甚至更优选为约6.5。优选将收集液保存在二氧化碳分压高于空气的大气中；例如，大气中二氧化碳分压可大于0.9，更优选0.95，还更优选0.99。
可将分选细胞收集至含有或包被有冰冻伸展剂的容器中，以代替常规收集液的使用。因此，在具体的实施方案中，分选细胞收集于含有运动性抑制剂的冰冻伸展剂中。在另一个实施方案中，分选细胞收集于这样的冰冻伸展剂中，其含有运动性抑制剂、水、
Figure BDA0000137094600000181
(Minitube，Verona，WI，含有甘油、tris、柠檬酸、果糖、泰乐菌素5mg/100ml、庆大霉素25mg/100ml、壮观霉素30mg/l00ml及林可霉素15mg/100ml)、卵黄以及丙酮酸盐。而在另一个实施方案中，收集液为这样的冰冻伸展剂，其含有0.097mol/LNaHCO3、0.173mol/L KHCO3、0.090mol/L C6H8O7·H2O的水溶液，且每75ml水中含25g
Figure BDA0000137094600000182
25g卵黄和10mM丙酮酸盐。
分选细胞的冰冻伸展
一旦完成精子的分选并收集其至收集容器内，这些精子即可用于对雌性哺乳动物授精。这几乎不需要对精子进行其它处理就可以立即进行。同样，也可以将精子冷却或冰冻以备后用。在这种情况下，加入冰冻伸展剂以最大程度地降低冷却或冰冻对存活力或解冻后运动性的影响，对精子大有益处。
运动性抑制剂可用于使冰冻伸展剂中的细胞不运动。一般而言，冰冻伸展剂含有运动性抑制剂、蛋白质源和冰冻保护剂。若含有，也可加入例如蛋白质源以提供细胞支持、缓冲细胞与收集容器的接触。蛋白质源可为任何不会干扰精子细胞存活力并与具体所用运动性抑制剂相容的蛋白质源。常用蛋白质源的实例包括乳(包括热匀浆和脱脂乳)、乳提取物、卵黄、卵黄提取物、大豆蛋白和大豆蛋白提取物。此类蛋白质的使用浓度可为约10％(v/v)至约30％(v/v)，优选约10％(v/v)至约20％(v/v)，更优选约20％(v/v)。
冰冻伸展剂中优选含有冰冻保护剂，以减轻或避免冷冻休克或以保持精子的生育力。本领域已知众多冰冻保护剂。适于与给定伸展剂共同使用的冰冻保护剂可有多种选择，取决于待冰冻的精子来源于哪一种系。适宜的冰冻保护剂的实例包括如，甘油、二甲基亚砜、乙二醇、丙二醇、海藻糖、
Figure BDA0000137094600000191
以及它们的组合。若含有冰冻保护剂，则其在冰冻伸展剂中的量一般为约1％(v/v)至约15％(v/v)，优选其量为约5％(v/v)至约10％(v/v)，更优选其量为约7％(v/v)，最优选量为约6％(v/v)。
在具体实施方案中，冰冻伸展剂包含运动性抑制剂、水、
Figure BDA0000137094600000192
卵黄和丙酮酸盐。而在另一个实施方案中，冰冻伸展剂为含0.097mol/LNaHCO3、0.173mol/L KHCO3、0.090mol/L C6H8O7·H2O的水溶液，且每75ml水中含25g25g卵黄和10mM丙酮酸盐。
在另一个具体实施方案中，冰冻伸展剂包含运动性抑制剂、水、
Figure BDA0000137094600000194
和卵黄。而在另一个实施方案中，冰冻伸展剂为含0.097mol/LNaHCO3、0.173mol/L KHCO3、0.090mol/L C6H8O7·H2O的水溶液，且每75ml水中含25g25g卵黄。
冰冻伸展剂中也可任选地含有如上文“细胞样品的收集”中讨论过的抗生素、生长因子或调节细胞内和/或细胞外氧化/还原反应的组合物。可据此将这些添加剂中的每一种加入收集液中。
上文已经详细描述了本发明，显而易见的是，在不脱离所附权利要求定义的本发明范围时可以对之进行修饰和改变。
实施例
提供以下非限制性实施例以进一步举例说明本发明。
实施例1
利用人工阴道从性成熟公牛中收集公牛精液，并将样品在25℃温控容器内转运至染色设施。精液一经接收，便根据标准和公知的方法(Farrell等人Theriogenology，49(4)：871-9(1998年3月))，利用Hamilton-Thorn运动性分析仪(IVOS)分析其浓度、运动性和前向运动性。根据精液浓度，将精液悬浮于TCA缓冲物或基于碳酸盐的抑制剂中，制备数管含150×106精子/ml的悬浮液。下表I显示了所用的组合物和染色条件。
表I
向精子悬浮液中加入等份10mM Hoechst水溶液以使浓度为600μMHoechst。染色期间将精子悬浮液留置于28℃水浴中(约1小时)。从染色分精子悬浮液中移取50μl小份，加入200μl 25℃含10mM丙酮酸盐的TCA(pH7.3)以起始静止逆转，至少5分钟的平衡时间后，通过IVOS分析测量前向性运动百分比(前向性运动％)进行分析。IVOS前向性运动百分比的比较结果见图1-3。
实施例2
利用人工阴道从性成熟公牛中收集公牛精液，并将样品在25℃温控容器内转运至染色设施。精液一经接收，便根据标准和公知的方法(Farrell等人Theriogenology，49(4)：871-9(1998年3月))，利用Hamilton-Thorn运动性分析仪(IVOS)分析其浓度、运动性和前向运动性。根据精液浓度，将精液悬浮于TCA缓冲物或基于碳酸盐的抑制剂中，制备数管含450×106精子/ml的悬浮液。下表II显示了所用的组合物和染色条件。
表II
Figure BDA0000137094600000211
向精子悬浮液中加入等份10mM Hoechst水溶液以使浓度为1000μMHoechst。将精子悬浮液留置于28℃水浴中1小时，然后按照各图中具体的说明，用含10mM丙酮酸盐的TCA或pH 6.2基于碳酸盐的抑制剂将之稀释为150×106精子/ml的典型分选浓度。在各图指定的时间内从染色并稀释的精子悬浮液中移取50μl小份，并加入200μl 25℃含10mM丙酮酸盐的TCA(pH7.3)以起始静止逆转，至少5分钟的平衡时间后，通过IVOS分析测量前向性运动百分比(前向性运动％)进行分析。IVOS前向性运动百分比的比较结果见图4-6。
实施例3
利用人工阴道从性成熟公牛中收集公牛精液，并将样品在25℃温控容器内转运至染色设施。精液一经接收，便根据标准和公知的方法(Farrell等人Theriogenology，49(4)：871-9(1998年3月))，利用Hamilton-Thorn运动性分析仪(IVOS)分析其浓度、运动性和前向运动性。根据精液浓度，将精液悬浮于TCA缓冲物或基于碳酸盐的抑制剂中，制备数管含450×106精子/ml的悬浮液。下表III显示了所用的组合物和染色条件。
表III
向精子悬浮液中加入等份10mM Hoechst水溶液以使浓度为300lμMHoechst。将精子悬浮液留置于41℃水浴中30分钟，然后按照各图中具体的说明，用含10mM丙酮酸盐的TCA或pH 6.2基于碳酸盐的抑制剂将之稀释为150×106精子/ml的典型分选浓度。在各图指定的时间内从染色并稀释的精子悬浮液中移取50μl小份，并加入200μl 25℃含10mM丙酮酸盐的TCA(pH7.3)以起始静止逆转，至少5分钟的平衡时间后，通过IVOS分析测量前向性运动百分比(前向性运动％)进行分析。IVOS前向性运动百分比的比较结果见图7-9。
实施例4
利用人工阴道从性成熟公牛中收集公牛精液，并将样品以2份的碳酸盐缓冲液稀释、于25℃温控容器内转运至染色设施。精液一经接收，便根据标准和公知的方法(Farrell等人Theriogenology，49(4)：871-9(1998年3月))，利用Hamilton-Thorn运动性分析仪(IVOS)分析其浓度、运动性和前向运动性。根据精液浓度，移取小份碳酸盐精子悬浮液、将其在500Xg离心5分钟、除去上清液并将沉淀重悬于41℃ pH 7.3的TCA缓冲液中，制备1mL 150×106精子/ml的悬浮液。再通过将小份精液悬浮于41℃含10mM丙酮酸盐、pH 7.3的TCA缓冲液内，制备另外1mL 150×106精子/ml悬浮液。向精子悬浮液中加入等份10mM Hoechst水溶液以使染料浓度为400μM Hoechst。染色期间将精子悬浮液留置于41℃水浴中。从染色的精子悬浮液中移取50μl小份，加入200μl相同温度的相同缓冲物，通过IVOS分析测量前向性运动百分比(前向性运动％)进行分析。IVOS分析的结果总结于图10。
实施例5
除以下不同外，按照实施例4的相同方法获取和制备精子样品：用于悬浮精子以进行染色和IVOS分析的缓冲物为TCA和含10μM维生素K的TCA。IVOS分析的结果总结于图11。
实施例6
除以下不同外，按照实施例4的相同方法获取和制备精子样品：用于悬浮精子以进行染色和IVOS分析的缓冲物为TCA和含100μM维生素K的TCA。IVOS分析的结果总结于图12。
实施例7
除以下不同外，按照实施例4的相同方法获取和制备精子样品：用于悬浮精子以进行染色和IVOS分析的缓冲物为TCA和含1mM硫辛酸的TCA。IVOS分析的结果总结于图13。
实施例8
利用人工阴道从性成熟公牛中收集公牛精液，并将样品以2份的碳酸盐缓冲液稀释、于25℃温控容器内转运至染色设施。精液一经接收，便根据标准和公知的方法(Farrell等人Theriogenology，49(4)：871-9(1998年3月))，利用Hamilton-Thorn运动性分析仪(IVOS)分析其浓度、运动性和前向运动性。根据精液浓度，将精子悬浮液在500Xg离心5分钟、除去上清液并将沉淀重悬于28℃ pH 7.3的TCA缓冲液中，制备1mL含150×106精子/ml的悬浮液。再通过将小份精液悬浮于28℃含10mM丙酮酸盐、pH 7.3的TCA缓冲液内，制备另外1mL 150×106精子/ml悬浮液。向精子悬浮液中加入等份10mM Hoechst水溶液以使染料浓度为600μMHoechst。染色期间将精子悬浮液留置于28℃水浴中。从染色精子悬浮液中移取50μl小份，加入200μl相同温度的相同缓冲物，通过IVOS分析测量前向性运动百分比(前向性运动％)进行分析。IVOS分析的结果总结于图14。
实施例9
除以下不同外，按照实施例8的相同方法获取和制备精子样品：用于悬浮精子以进行染色和IVOS分析的缓冲物为TCA和含100μM维生素K的TCA。IVOS分析的结果总结于图15。
实施例10
除以下不同外，按照实施例8的相同方法获取和制备精子样品：用于悬浮精子以进行染色和IVOS分析的缓冲物为TCA和含1mM硫辛酸的TCA。IVOS分析的结果总结于图16。
实施例11
利用人工阴道从性成熟公牛中收集公牛精液，并将样品以2份的碳酸盐缓冲液稀释、于25℃温控容器内转运至染色设施。精液一经接收，便根据标准和公知的方法(Farrell等人Theriogenology，49(4)：871-9(1998年3月))，利用Hamilton-Thorn运动性分析仪(IVOS)分析其浓度、运动性和前向运动性。根据精液浓度，移取小份碳酸盐精子悬浮液、将其在500Xg离心5分钟、除去上清液并将沉淀重悬于1ml TCA缓冲液或1ml含2.5mM、10mM、25mM或50mM丙酮酸盐的TCA缓冲液中，制备1mL含150×106精子/ml的悬浮液。向样品中加入MON33342溶液以使染料终浓度为600μM。于28℃水浴中孵育悬浮液。从染色精子悬浮液中移取50μl小份，加入200μl相同温度的相同缓冲物，通过IVOS分析测量前向性运动百分比(前向性运动％)进行分析。IVOS分析前向性运动％的结果如图17所示。
实施例12
利用人工阴道从性成熟公牛中收集公牛精液，并将样品以2份的碳酸盐缓冲液稀释、于25℃温控容器内转运至染色设施。精液一经接收，便根据标准和公知的方法(Farrell等人Theriogenology，49(4)：871-9(1998年3月))，利用Hamilton-Thorn运动性分析仪(IVOS)分析其浓度、运动性和前向运动性。根据精液浓度，移取小份碳酸盐精子悬浮液、将其在500Xg离心5分钟、除去上清液并将沉淀重悬于1ml TCA缓冲液中，制备1mL含150×106精子/ml的TCA缓冲液。移取一份碳酸盐精子悬浮液、将其在500Xg中离心5分钟、除去上清液并将沉淀重悬于1ml含10mM丙酮酸盐的TCA缓冲液中，制备1mL含150×106精子/ml的10mM丙酮酸盐TCA缓冲液。向样品中加入SYBR14染料溶液以使染料终浓度为20μM。于28℃水浴中孵育悬浮液。从染色精子悬浮液中移取50μl小份，加入200μl相同温度的相同缓冲物，通过IVOS分析测量前向性运动百分比(前向性运动％)进行分析。IVOS分析前向性运动％的结果如图18所示。
实施例13
利用人工阴道从性成熟公牛中收集公牛精液，并将样品以2份的碳酸盐缓冲液稀释、以于25℃温控容器内转运至染色设施。精液一经接收，便根据标准和公知的方法(Farrell等人Theriogenology，49(4)：871-9(1998年3月))，利用Hamilton-Thorn运动性分析仪(IVOS)分析其浓度、运动性和前向运动性。根据精液浓度，移取小份碳酸盐精子悬浮液、将其在500Xg离心5分钟、除去上清液并将沉淀重悬于1ml TCA缓冲液中，制备1mL含150×106精子/ml的TCA缓冲液。移取一份碳酸盐精子悬浮液、将其在500Xg中离心5分钟、除去上清液并重悬浮沉淀于1ml含10mM丙酮酸盐的TCA缓冲液中，制备1mL含150×106精子/ml的10mM丙酮酸盐TCA缓冲液。向样品中加入BBC溶液以使染料终浓度为100μM。于28℃水浴中孵育悬浮液。从染色精子悬浮液中移取50μl小份，加入200μl相同温度的相同缓冲物，通过IVOS分析测量前向性运动百分比(前向性运动％)进行分析。IVOS分析前向性运动％的结果如图19所示。
实施例14
除以下不同外，按照实施例4的相同方法获取和制备精子样品：染色浓度为200μM BBC。IVOS分析的结果总结于图20。

Claims (20)

1.制备性别富集的精子细胞悬浮液的方法，包括将精子细胞悬浮液与抑制运动性的组合物混合以形成化学不运动的精子细胞悬浮液，用DNA选择性染料染色所述化学不运动的精子细胞悬浮液，以及分选染色的化学不运动的精子细胞悬浮液以形成带有X染色体或带有Y染色体的精子细胞的活的、性别富集的授精群，所述精子细胞具有与其DNA相结合的DNA选择性染料。
2.权利要求1的方法，其中染料从由Hoechst 33342、Hoechst 33258、SYBR-14、以及双苯甲亚胺-
Figure FDA0000137094590000011
缀合物6-{[3-((2Z)-2-{[1-(二氟硼烷基)-3，5-二甲基-1H-吡咯-2-基]亚甲基}-2H-吡咯-5-基)丙酰基]氨基}-N-[3-(甲基{3-[({4-[6-(4-甲基哌嗪-1-基)-1H，3’H-2，5’-双苯并咪唑-2’-基]苯氧基}乙酰基)氨基]丙基}氨基)丙基]己酰胺组成的组中选择。
3.权利要求1的方法，其中在分选方法中通过抑制运动性的含有碳酸盐源的组合物使得化学不运动的精子细胞悬浮液化学不运动。
4.权利要求3的方法，其中在分选方法中通过抑制运动性的含有NaHCO3、KHCO3和C6H8O7·H2O的组合物使得所述化学不运动的精子细胞悬浮液化学不运动。
5.权利要求4的方法，其中在分选方法中通过抑制运动性的含有水中0.097mol/L NaHCO3、0.173mol/L KHCO3、0.090mol/L C6H8O7·H2O的组合物使得所述化学不运动的精子细胞悬浮液化学不运动。
6.权利要求1的方法，其中在悬浮液中所述带有X染色体的精子细胞或带有Y染色体的精子细胞的活的、性别富集的授精群的精子浓度为至少1.25×108精子/ml。
7.权利要求1的方法，其中在悬浮液中带有X染色体的精子细胞或带有Y染色体的精子细胞的所述活的、性别富集的授精群的精子浓度为至少1.5×108精子/ml。
8.权利要求1的方法，其中在悬浮液中带有X染色体的精子细胞或带有Y染色体的精子细胞的所述活的、性别富集的授精群的精子浓度为至少1.75×108精子/ml。
9.权利要求1的方法，其中在悬浮液中带有X染色体的精子细胞或带有Y染色体的精子细胞的所述活的、性别富集的授精群的精子浓度为小于约9.0×105精子/ml。
10.权利要求1的方法，其中在悬浮液中带有X染色体的精子细胞或带有Y染色体的精子细胞的所述活的、性别富集的授精群的精子浓度为小于约7×105精子/ml。
11.权利要求1的方法，其中在悬浮液中带有X染色体的精子细胞或带有Y染色体的精子细胞的所述活的、性别富集的授精群的精子浓度为小于约5×105精子/ml。
12.权利要求1的方法，所述在悬浮液中带有X染色体的精子细胞或带有Y染色体的精子细胞的所述活的、性别富集的授精群的精子浓度为小于约2×105精子/ml。
13.权利要求1的方法，其中在悬浮液中带有X染色体的精子细胞或带有Y染色体的精子细胞的所述活的、性别富集的授精群的精子浓度为小于约1×105精子/ml。
14.权利要求1的方法，其中所述活的精子悬浮液包含活的冷冻的精子。
15.权利要求1的方法，其中所述带有X染色体的精子细胞或带有Y染色体的精子细胞的活的、性别富集的授精群的浓度是小于约1×106精子/ml或至少为约1×108精子/ml的在悬浮液中的精子。
16.权利要求1的方法，其中在分选方法中在包含钾和钠的悬浮液中使得化学不运动的精子细胞悬浮液化学不运动，其中钾与钠的摩尔比大于各1∶1。
17.权利要求16的方法，其中钾与钠的摩尔比大于各1.25∶1。
18.权利要求16的方法，其中钾与钠的摩尔比大于各1.5∶1。
19.权利要求16的方法，其中钾与钠的摩尔比大于各1.75∶1。
20.权利要求16的方法，其中钾与钠的摩尔比大于各2∶1。
CN201210040778.6A 2004-03-29 2005-03-29 用于分选带有x或y染色体富集群的精子悬浮液 Active CN102586179B (zh)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US55740704P 2004-03-29 2004-03-29
US60/557,407 2004-03-29
US61417804P 2004-09-29 2004-09-29
US60/614,178 2004-09-29
US61844004P 2004-10-13 2004-10-13
US60/618,440 2004-10-13

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN2005800173705A Division CN101014700B (zh) 2004-03-29 2005-03-29 用于分选带有x或y染色体富集群的精子悬浮液

Publications (2)

Publication Number Publication Date
CN102586179A true CN102586179A (zh) 2012-07-18
CN102586179B CN102586179B (zh) 2015-04-08

Family

ID=34964337

Family Applications (2)

Application Number Title Priority Date Filing Date
CN201510400906.7A Pending CN104974983A (zh) 2004-03-29 2005-03-29 用于授精的精子悬浮液
CN201210040778.6A Active CN102586179B (zh) 2004-03-29 2005-03-29 用于分选带有x或y染色体富集群的精子悬浮液

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN201510400906.7A Pending CN104974983A (zh) 2004-03-29 2005-03-29 用于授精的精子悬浮液

Country Status (16)

Country Link
US (2) US7892725B2 (zh)
EP (5) EP2801363B1 (zh)
JP (4) JP2007537727A (zh)
CN (2) CN104974983A (zh)
AR (2) AR049259A1 (zh)
AU (2) AU2005228893B2 (zh)
BR (2) BRPI0509485A (zh)
CA (3) CA2972254A1 (zh)
DK (2) DK2801363T3 (zh)
ES (1) ES2397678T3 (zh)
HK (2) HK1173745A1 (zh)
MX (2) MXPA06011345A (zh)
NZ (2) NZ550198A (zh)
PL (1) PL2151243T3 (zh)
PT (1) PT2151243E (zh)
WO (2) WO2005094852A2 (zh)

Families Citing this family (47)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2279574C (en) 1997-01-31 2007-07-24 The Horticulture & Food Research Institute Of New Zealand Ltd. Optical apparatus
US6149867A (en) 1997-12-31 2000-11-21 Xy, Inc. Sheath fluids and collection systems for sex-specific cytometer sorting of sperm
NZ509434A (en) 1998-07-30 2004-03-26 Univ Colorado State Res Found Equine system for non-surgical artificial insemination
US7208265B1 (en) 1999-11-24 2007-04-24 Xy, Inc. Method of cryopreserving selected sperm cells
WO2001085913A2 (en) 2000-05-09 2001-11-15 Xy, Inc. High purity x-chromosome bearing and y-chromosome bearing populations of spermatozoa
US7713687B2 (en) 2000-11-29 2010-05-11 Xy, Inc. System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations
CA2468774C (en) 2000-11-29 2015-06-30 George E. Seidel System for in-vitro fertilization with spermatozoa separated into x-chromosome and y-chromosome bearing populations
US8486618B2 (en) 2002-08-01 2013-07-16 Xy, Llc Heterogeneous inseminate system
EP2275533B9 (en) 2002-08-01 2016-10-19 Xy, Llc Method of assessing sperm cells
US7855078B2 (en) 2002-08-15 2010-12-21 Xy, Llc High resolution flow cytometer
US7169548B2 (en) 2002-09-13 2007-01-30 Xy, Inc. Sperm cell processing and preservation systems
DK2959774T3 (da) 2003-03-28 2019-05-13 Inguran Llc Fremgangsmåde til kryokonservering af sædceller
WO2004104178A2 (en) * 2003-05-15 2004-12-02 Xy, Inc. Efficient haploid cell sorting for flow cytometer systems
BRPI0509485A (pt) 2004-03-29 2007-09-11 Monsanto Technology Llc suspensões de esperma para uso em inseminação
EP1769067A2 (en) 2004-07-22 2007-04-04 Monsanto Technology LLC Process for enriching a population of sperm cells
DE102005044530B4 (de) * 2004-09-16 2010-02-18 Masterrind Gmbh Verfahren zur geschlechtsspezifischen Selektion von Säugerspermatozoen
US9433484B2 (en) * 2007-07-27 2016-09-06 Brad K. Stroud Artificial breeding techniques for bovines including semen diluents and AI apparatus
US8415094B2 (en) * 2007-12-21 2013-04-09 Jaffar Ali bin M. Abdullah Protein-free gamete and embryo handling and culture media products
CN101554152B (zh) * 2009-05-21 2012-05-09 西北农林科技大学 一种冷冻精液稀释液及其配制方法
EP2503956A4 (en) * 2009-11-24 2013-06-26 Biassex Pty Ltd METHOD FOR INFLUENCING SEXUAL SELECTION IN ARTIFICIAL INSEMINATION AND APPARATUS THEREFOR
US10610343B2 (en) 2013-07-03 2020-04-07 Brad K. Stroud Method, apparatus and kit for artificial insemination of bovine
US9554883B2 (en) 2010-08-10 2017-01-31 Brad K. Stroud Method and apparatus to reduce the number of sperm used in artificial insemination of cattle
US11622844B2 (en) 2010-08-10 2023-04-11 Maximate, Llc Method, apparatus and kit for artificial insemination of bovine
US9340762B2 (en) 2010-08-20 2016-05-17 Yu Sun Method for automated sperm manipulation and device for holding sperm and oocytes
EP2713713A4 (en) 2011-06-01 2014-12-10 Inguran Llc COMPOSITIONS AND METHODS FOR IMPROVING THE QUALITY OF PROCESSED SPERM
US9781919B2 (en) 2011-06-01 2017-10-10 Inguran, Llc Compositions and methods for improving the quality of processed sperm
LT2768607T (lt) 2011-09-26 2021-12-27 Thermo Fisher Scientific Geneart Gmbh Daugiašulininė plokštelė didelio efektyvumo, mažo tūrio nukleorūgščių sintezei atlikti
BR112014022817A8 (pt) 2012-03-14 2021-07-13 Membrane Protective Tech Inc processo de aumento da criopreservação de células biológicas e composição de criopreservação
WO2013152220A2 (en) 2012-04-04 2013-10-10 Life Technologies Corporation Tal-effector assembly platform, customized services, kits and assays
US9888990B2 (en) 2012-06-06 2018-02-13 Inguran, Llc Methods for use of sex sorted semen to improve genetic management in swine
US9433195B2 (en) 2012-06-06 2016-09-06 Inguran, Llc Methods for increasing genetic progress in a line or breed of swine using sex-selected sperm cells
US9034161B2 (en) 2012-08-23 2015-05-19 Douglas T. Carrell Sperm separation devices and associated methods
CA2886782C (en) * 2012-10-05 2020-03-10 Inguran, Llc High efficiency methods of sex sorting sperm
US10620213B2 (en) 2012-10-05 2020-04-14 Inguran, Llc High pressure sperm sorting and flow cytometer methods
US10563240B2 (en) 2013-03-14 2020-02-18 Life Technologies Corporation High efficiency, small volume nucleic acid synthesis
US20160017394A1 (en) 2014-07-15 2016-01-21 Life Technologies Corporation Compositions and methods for nucleic acid assembly
JP6832848B2 (ja) * 2014-10-20 2021-02-24 ザ ユニバーシティ オブ ユタ リサーチ ファウンデイション 組織試料処理システム及びそれに関連する方法
CA3210347A1 (en) 2014-11-20 2016-05-26 Inguran, Llc Low sugar sperm media and compositions
LT3557262T (lt) 2014-12-09 2022-11-10 Life Technologies Corporation Didelio efektyvumo nukleorūgščių sintezė mažame tūryje
WO2017007925A1 (en) 2015-07-07 2017-01-12 Thermo Fisher Scientific Geneart Gmbh Enzymatic synthesis of nucleic acid sequences
EP3359669B1 (en) 2015-10-06 2020-05-13 Pierce Biotechnology, Inc. Devices and methods for producing proteins
PT110231B (pt) * 2017-08-02 2021-02-22 Universidade Do Porto Potenciadores de mtor e suas utilizações para melhorar a qualidade e função do esperma durante o armazenamento
EP3814494B1 (en) 2018-06-29 2023-11-01 Thermo Fisher Scientific GENEART GmbH High throughput assembly of nucleic acid molecules
US10470798B1 (en) 2018-11-30 2019-11-12 Ohana Biosciences, Inc. Methods for promoting fertilization
GB201905303D0 (en) 2019-04-15 2019-05-29 Thermo Fisher Scient Geneart Gmbh Multiplex assembly of nucleic acid molecules
EP4114972A1 (en) 2020-03-06 2023-01-11 Life Technologies Corporation High sequence fidelity nucleic acid synthesis and assembly
DE102021002257A1 (de) 2021-04-29 2022-11-03 Forschungsverbund Berlin E.V. Mittel zur Konservierung von Säugetiersperma und Verwendung des Mittels

Family Cites Families (635)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3005756A (en) * 1958-11-14 1961-10-24 Noland L Van Demark Diluter containing carbon dioxide for preserving semen
US3005759A (en) * 1959-04-24 1961-10-24 American Zinc Inst Zinc electroplating
US3299354A (en) 1962-07-05 1967-01-17 Coulter Electronics Aperture tube structure for particle study apparatus
US3499435A (en) 1967-06-02 1970-03-10 Paul E Rockwell Esophageal probe for use in monitoring
US3547526A (en) 1967-10-26 1970-12-15 Kollsman Instr Corp Optical beam cross-section converter
US3810010A (en) 1968-11-02 1974-05-07 Telefunken Patent Particle analysis method and apparatus wherein liquid containing particles is sucked into a constricted flow path
US3829216A (en) 1968-11-26 1974-08-13 M Persidsky Optical system and method for counting sperm cells
DE1815352C3 (de) 1968-12-18 1975-03-20 Wolfgang Prof. Dr. Dittrich Automatisches MeB- und Zählgerät für die Teilchen einer Dispersion
US3894529A (en) 1969-04-10 1975-07-15 Bio Controls Inc Method and means for controlling the sex of mammalian offspring and product therefor
US4327177A (en) 1969-04-10 1982-04-27 Wallace Shrimpton Method and means for controlling the sex of mammalian offspring and product therefor
US4474875A (en) * 1969-04-10 1984-10-02 Wallace Shrimpton Method and means for controlling the sex of mammalian offspring and product therefor
DE1919628C3 (de) 1969-04-18 1975-04-10 Wolfgang Prof. Dr. Dittrich Anordnung zum automatischen Zählen und/oder Klassifizieren von in einem strömungsfähigen Medium dispergierten Teilchen
US3687806A (en) 1969-11-04 1972-08-29 Bio Controls Inc Method for controlling sex of mammalian offspring
US3788744A (en) 1970-01-14 1974-01-29 Bio Physics Systems Inc Method and apparatus for photoanalysis
US3661460A (en) 1970-08-28 1972-05-09 Technicon Instr Method and apparatus for optical analysis of the contents of a sheathed stream
US3816249A (en) 1970-11-23 1974-06-11 B Bhattacharya Universal medium and method for extending the useful life of semen in vitro
US3644128A (en) 1970-12-28 1972-02-22 Stuart Lipner Method of preparing comminuted meat products
US3756459A (en) 1971-01-12 1973-09-04 Damon Corp Method and apparatus for metering fluid utilizing pressure differentials
US3791384A (en) 1971-07-15 1974-02-12 Schaumann H Artificial insemination of sows
US3833796A (en) 1971-10-13 1974-09-03 Georgia Tech Res Inst Method and apparatus for chromosome digitizing
BE793185A (fr) 1971-12-23 1973-04-16 Atomic Energy Commission Appareil pour analyser et trier rapidement des particules telles que des cellules biologiques
US3826364A (en) 1972-05-22 1974-07-30 Univ Leland Stanford Junior Particle sorting method and apparatus
US3761941A (en) 1972-10-13 1973-09-25 Mead Corp Phase control for a drop generating and charging system
CA1029833A (en) 1973-02-23 1978-04-18 Hildegarde Goehde Apparatus for the automatic counting and measuring of suspended particles
US3791517A (en) 1973-03-05 1974-02-12 Bio Physics Systems Inc Digital fluidic amplifier particle sorter
US4009260A (en) 1973-04-19 1977-02-22 Schering Aktiengesellschaft Fractionation of sperm
USRE29141E (en) 1973-06-14 1977-02-22 Coulter Electronics, Inc. Apparatus for orienting generally flat particles for sensing
US3893766A (en) 1973-06-14 1975-07-08 Coulter Electronics Apparatus for orienting generally flat particles for slit-scan photometry
US3947093A (en) 1973-06-28 1976-03-30 Canon Kabushiki Kaisha Optical device for producing a minute light beam
US3944917A (en) 1973-08-13 1976-03-16 Coulter Electronics, Inc. Electrical sensing circuitry for particle analyzing device
US3909744A (en) 1973-09-24 1975-09-30 United Technologies Corp Unstable resonator system producing a high irradiance beam in the far field
US3906929A (en) 1973-11-23 1975-09-23 Lynn Lawrence Augspurger Processes for reproduction of cellular bodies
FR2254639B1 (zh) 1973-12-18 1978-06-02 Agronomique Inst Nat Rech
US4070617A (en) 1974-05-08 1978-01-24 Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. Device for controlling the particle flow in an apparatus for measuring the properties of particles suspended in liquid
US3973196A (en) 1974-05-24 1976-08-03 Coulter Electronics, Inc. Method and apparatus for ejecting a metered amount of particulate sample
US3963606A (en) 1974-06-03 1976-06-15 Coulter Electronics, Inc. Semi-automatic adjusting delay for an electronic particle separator
US3877430A (en) 1974-07-17 1975-04-15 Horst K Wieder Artificial insemination apparatus
US4083957A (en) 1974-07-26 1978-04-11 Lang John L Process for the alteration of the sex-ratio of mammals
US4006360A (en) 1974-08-21 1977-02-01 Block Engineering, Inc. Method of discriminating between dyed particles and background fluorescence of the dye
US4092229A (en) 1975-12-17 1978-05-30 Bhattacharya Bhairab C Thermal convection counter streaming sedimentation and forced convection galvanization method for controlling the sex of mammalian offspring
US3976197A (en) 1974-11-22 1976-08-24 Bhattacharya Bhairab C Thermal convection counter streaming sedimentation method and apparatus for controlling the sex of mammalian offspring
US4014611A (en) 1975-04-30 1977-03-29 Coulter Electronics, Inc. Aperture module for use in particle testing apparatus
DE2521236C3 (de) 1975-05-10 1978-12-14 Hildegard Dr. 4400 Muenster Goehde Geb. Kuhl Einrichtung zum Zählen und Messen von in einer Flüssigkeit suspendierten Teilchen
US3960449A (en) 1975-06-05 1976-06-01 The Board Of Trustees Of Leland Stanford Junior University Measurement of angular dependence of scattered light in a flowing stream
US4058732A (en) 1975-06-30 1977-11-15 Analytical Radiation Corporation Method and apparatus for improved analytical fluorescent spectroscopy
US4007087A (en) 1975-10-17 1977-02-08 Gametrics Limited Sperm fractionation and storage
AU2154077A (en) 1976-01-27 1978-07-27 Univ Edinburgh Control of sex ratio in mammalian offspring
US4302166A (en) 1976-04-22 1981-11-24 Coulter Electronics, Inc. Droplet forming apparatus for use in producing uniform particles
US4162282A (en) 1976-04-22 1979-07-24 Coulter Electronics, Inc. Method for producing uniform particles
GB1563856A (en) 1976-06-10 1980-04-02 Coulter Electronics Methods and apparatus for delectively separating small particles suspended in a liquid
GB1583150A (en) 1976-08-02 1981-01-21 Milk Marketing Board Apparatus for collecting eggs
US4110604A (en) 1976-11-04 1978-08-29 Becton, Dickinson And Company Particle density measuring system
DE2709399C3 (de) 1977-03-04 1980-07-24 Goehde, Wolfgang, Dr., 4400 Muenster Einrichtung zum Messen von Zelleigenschaften
DE2716095A1 (de) 1977-04-12 1978-10-19 Zoeld Tibor Dr Phys Gasgesteuertes verfahren zum sortieren von in einem elektrolyten suspendierten teilchen und vorrichtung zur durchfuehrung des verfahrens
US4448767A (en) 1977-10-11 1984-05-15 Sumar Corporation Preparation of monospecific male-specific antibody and the use thereof for increasing the percentage of mammalian offspring of either sex
US4191749A (en) 1977-10-11 1980-03-04 Bryant Bernard J Method and material for increasing the percentage of mammalian offspring of either sex
US4189236A (en) 1978-03-20 1980-02-19 Coulter Electronics, Inc. Ellipsoid-conic radiation collector and method
US4179218A (en) 1978-05-15 1979-12-18 The Boeing Company Particle size analyzer
US4251733A (en) 1978-06-29 1981-02-17 Hirleman Jr Edwin D Technique for simultaneous particle size and velocity measurement
DE2832091A1 (de) 1978-07-21 1980-01-31 Eidenschink Henning Optisches verfahren zur bestimmung der teilchengroesse kolloidaler loesungen und messgeraet zur durchfuehrung des verfahrens
US4225405A (en) 1978-08-16 1980-09-30 Lawson Rommom L Process for separation and collection of viable female and male spermatozoa
US4276139A (en) 1978-08-16 1981-06-30 Lawson Rommon L Process for magnetic separation and collection of viable female and male spermatozoa
US4230558A (en) 1978-10-02 1980-10-28 Coulter Electronics, Inc. Single drop separator
US4341471A (en) 1979-01-02 1982-07-27 Coulter Electronics, Inc. Apparatus and method for measuring the distribution of radiant energy produced in particle investigating systems
US4267268A (en) 1979-03-12 1981-05-12 Nelson Jr Robert A Spermatozoa extenders
US4274408A (en) 1979-03-26 1981-06-23 Beatrice Nimrod Method for guide-wire placement and novel syringe therefor
US4200802A (en) 1979-03-28 1980-04-29 The United States Of America As Represented By The United States Department Of Energy Parabolic cell analyzer
US4408877A (en) 1979-04-10 1983-10-11 Ernst Leitz Wetzlar Gmbh Device for hydrodynamic focussing of a particle-suspension in a liquid flow cytophotometer
NO144002C (no) 1979-04-10 1981-05-27 Norsk Hydro S Inst For Kreftfo Anordning for anvendelse ved vaeskestroemsfotometri
US4263508A (en) 1979-04-20 1981-04-21 Research Corporation Pulse edge measurement for determining particle dimensional characteristics
US4255021A (en) 1979-04-20 1981-03-10 The United States Of America As Represented By The United States Department Of Energy Optical device with conical input and output prism faces
US4325483A (en) 1979-08-20 1982-04-20 Ortho Diagnostics, Inc. Method for detecting and controlling flow rates of the droplet forming stream of an electrostatic particle sorting apparatus
US4317520A (en) 1979-08-20 1982-03-02 Ortho Diagnostics, Inc. Servo system to control the spatial position of droplet formation of a fluid jet in a cell sorting apparatus
US4318480A (en) 1979-08-20 1982-03-09 Ortho Diagnostics, Inc. Method and apparatus for positioning the point of droplet formation in the jetting fluid of an electrostatic sorting device
US4318481A (en) 1979-08-20 1982-03-09 Ortho Diagnostics, Inc. Method for automatically setting the correct phase of the charge pulses in an electrostatic flow sorter
US4318482A (en) 1979-08-20 1982-03-09 Ortho Diagnostics, Inc. Method for measuring the velocity of a perturbed jetting fluid in an electrostatic particle sorting system
DE2943116C2 (de) 1979-10-25 1986-06-19 Gesellschaft für Strahlen- und Umweltforschung mbH, 8000 München Einrichtung zur durchflußcytometrischen Reaktions- und/oder Diffusionsmessung
US4284355A (en) 1979-10-29 1981-08-18 Ortho Diagnostics, Inc. Automated method for cell volume determination
US4400764A (en) 1980-02-05 1983-08-23 The Boeing Company Low backscatter illumination system
US4367043A (en) 1980-05-05 1983-01-04 Leland Stanford Junior University Method and means for delivering liquid samples to a sample scanning device
US4362246A (en) 1980-07-14 1982-12-07 Adair Edwin Lloyd Method of treating collected mammal semen and separating sperm into X Y components
US4348107A (en) 1980-07-18 1982-09-07 Coulter Electronics, Inc. Orifice inside optical element
EP0046345A3 (en) 1980-08-15 1982-03-03 Ortho Diagnostic Systems Inc. Controlled hydrodynamic flow in flow cytometry systems
US4350410A (en) 1980-10-08 1982-09-21 The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration Multiprism collimator
US4691829A (en) 1980-11-03 1987-09-08 Coulter Corporation Method of and apparatus for detecting change in the breakoff point in a droplet generation system
US4487320A (en) 1980-11-03 1984-12-11 Coulter Corporation Method of and apparatus for detecting change in the breakoff point in a droplet generation system
US4395676A (en) 1980-11-24 1983-07-26 Coulter Electronics, Inc. Focused aperture module
US4361400A (en) 1980-11-26 1982-11-30 The United States Of America As Represented By The United States Department Of Energy Fluidic assembly for an ultra-high-speed chromosome flow sorter
US4680258A (en) 1981-03-24 1987-07-14 Sloan-Kettering Institute For Cancer Research Process for sex determination in man by use of monoclonal antibodies to the H-Y antigen
US4673288A (en) 1981-05-15 1987-06-16 Ratcom, Inc. Flow cytometry
US4818103A (en) 1981-05-15 1989-04-04 Ratcom Flow cytometry
DE3266669D1 (en) 1981-06-24 1985-11-07 Becton Dickinson Co Analyzer for simultaneously determining volume and light emission characteristics of particles
US4339434A (en) 1981-08-17 1982-07-13 Gametrics Limited Method of increasing the incidence of female offspring
US4395397A (en) 1981-09-17 1983-07-26 Sidney Farber Cancer Institute, Inc. Apparatus and method for killing unwanted cells
US4422761A (en) 1981-09-28 1983-12-27 Frommer Joseph C Photo-electric particle sensing system
US4515274A (en) 1981-12-02 1985-05-07 Coulter Corporation Particle analyzing and sorting apparatus
JPS59500340A (ja) 1982-03-08 1984-03-01 モトロ−ラ・インコ−ポレ−テツド 集積回路のリ−ドフレ−ム
US4511661A (en) 1982-03-19 1985-04-16 University Patents, Inc. ATCC HB8116 And its monoclonal anti-H-Y antibody, Hyclonalan
US4498766A (en) 1982-03-25 1985-02-12 Becton, Dickinson And Company Light beam focal spot elongation in flow cytometry devices
DE3315195A1 (de) 1982-04-29 1983-11-03 International Remote Imaging Systems Inc., 91311 Chatsworth, Calif. Verfahren zum ausrichten von teilchen in einer fluidprobe
DE3315194A1 (de) 1982-04-29 1983-11-03 International Remote Imaging Systems Inc., 91311 Chatsworth, Calif. Verfahren zum trennen von in einer fluidprobe stroemenden teilchen
US4629687A (en) 1982-07-29 1986-12-16 Board Of Trustees Of Michigan State University Positive selection sorting of cells
GB2125181B (en) 1982-08-11 1986-01-29 Coulter Electronics Flow cells for particle study
US4559309A (en) 1982-09-01 1985-12-17 Memorial Sloan Kettering Cancer Center Flow cytometry-fluorescence measurements for characterizing sperm
WO1984001265A1 (en) 1982-10-05 1984-04-12 Genetic Engineering Inc Method of treating collected mammal semen and separating sperm into x and y components
US4501366A (en) 1982-12-14 1985-02-26 Adolph Coors Company Photomultiplier tube assembly
DE3372137D1 (en) 1982-12-21 1987-07-23 Crosfield Electronics Ltd Light beam-splitter
US4492436A (en) 1983-01-03 1985-01-08 At&T Bell Laboratories Polarization independent beam splitter
JPS59143146A (ja) 1983-02-07 1984-08-16 Nippon Kogaku Kk <Nikon> ミラ−集光型照明光学系
CA1206559A (en) 1983-03-04 1986-06-24 Robert E. Auer Method of and apparatus for detecting change in the breakoff point of a droplet generation system
IT1197570B (it) 1983-02-11 1988-12-06 Serono Ist Farm Miscele di fsh e lh da ipofisi porcine in rapporto definito
CH651930A5 (en) 1983-03-24 1985-10-15 Coulter Corp Apparatus for analysis and sorting of particles
JPS59174742A (ja) 1983-03-25 1984-10-03 Agency Of Ind Science & Technol 微小粒子を区分又は選別する方法及びその装置
GB2145112B (en) 1983-04-27 1987-02-18 Milk Marketing Board Sorting living spermatozoa
US4523809A (en) 1983-08-04 1985-06-18 The United States Of America As Represented By The Secretary Of The Air Force Method and apparatus for generating a structured light beam array
NZ207393A (en) 1983-08-05 1987-03-31 Neal Lloyd First Staining dna in living cells
US4538733A (en) 1983-10-14 1985-09-03 Becton, Dickinson And Company Particle sorter with neutralized collection wells and method of using same
US4585736A (en) 1983-10-18 1986-04-29 The United States Of America As Represented By The United States Department Of Energy Flow cytometric measurement of total DNA and incorporated halodeoxyuridine
US4780406A (en) 1983-10-18 1988-10-25 The Regents Of The University Of California Flow cytometric measurement of total DNA and incorporated halodeoxyuridine
US4735504A (en) 1983-10-31 1988-04-05 Technicon Instruments Corporation Method and apparatus for determining the volume & index of refraction of particles
ATE58017T1 (de) 1983-12-24 1990-11-15 Inotech Ag Vorrichtung zum fuehren und sammeln von licht in der fotometrie od. dgl.
US4573796A (en) 1984-01-06 1986-03-04 The United States Of America As Represented By The United States Department Of Energy Apparatus for eliminating background interference in fluorescence measurements
US4631483A (en) 1984-02-01 1986-12-23 Coulter Electronics, Inc. Particle analyzing apparatus and method of moving particles in suspension through such apparatus
US4965204A (en) 1984-02-06 1990-10-23 The Johns Hopkins University Human stem cells and monoclonal antibodies
US4714680B1 (en) 1984-02-06 1995-06-27 Univ Johns Hopkins Human stem cells
WO1985004014A1 (en) 1984-02-29 1985-09-12 Research Corporation Flow cytometers
US4545677A (en) 1984-03-05 1985-10-08 Becton, Dickinson And Company Prismatic beam expander for light beam shaping in a flow cytometry apparatus
US4600302A (en) 1984-03-26 1986-07-15 Becton, Dickinson And Company Flow cytometry apparatus with uniform incoherent light excitation
DE3412620A1 (de) 1984-04-04 1985-10-17 Basf Ag, 6700 Ludwigshafen Laseroptische anordnung zur messung des dispergiergrades in stroemenden systemen
US4609286A (en) 1984-04-16 1986-09-02 Becton, Dickinson And Company Dispersion prism for separation of wavelengths of spectrally rich light in a flow cytometry apparatus
US4605558A (en) 1984-04-20 1986-08-12 Wallace Shrimpton Process for cell separation
US4660971A (en) 1984-05-03 1987-04-28 Becton, Dickinson And Company Optical features of flow cytometry apparatus
FR2563726B1 (fr) 1984-05-04 1986-10-10 Robert Cassou Appareil d'insemination artificielle, notamment des carnivores
FR2566543B1 (fr) 1984-06-20 1988-02-26 Commissariat Energie Atomique Dispositif optique a rendement de collection eleve et cytofluorimetre en faisant application
EP0171676B1 (en) 1984-07-31 1990-11-07 Hitachi, Ltd. Free-flow electrophoretic separation method and apparatus therefor
US4661913A (en) 1984-09-11 1987-04-28 Becton, Dickinson And Company Apparatus and method for the detection and classification of articles using flow cytometry techniques
DE3574617D1 (de) 1984-09-11 1990-01-11 Partec Ag Verfahren und vorrichtung zur sortierung von mikroskopischen partikeln.
US4598408A (en) 1984-10-22 1986-07-01 Trw Inc. High extraction efficiency cylindrical ring resonator
JPS61139747A (ja) 1984-12-12 1986-06-27 Canon Inc 粒子解析装置
JPS61159135A (ja) 1984-12-31 1986-07-18 Canon Inc 粒子解析装置
US4702598A (en) 1985-02-25 1987-10-27 Research Corporation Flow cytometer
US4683195A (en) 1986-01-30 1987-07-28 Cetus Corporation Process for amplifying, detecting, and/or-cloning nucleic acid sequences
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
CA1250808A (en) 1985-04-29 1989-03-07 David W. Dresser Semen sexing
US4662742A (en) 1985-05-10 1987-05-05 Becton, Dickinson And Company Scatter/fluorescene beam splitter in a flow cytometry apparatus
US4877965A (en) 1985-07-01 1989-10-31 Diatron Corporation Fluorometer
USRE34782E (en) 1985-07-01 1994-11-08 Diatron Corporation Fluorometer
US4744090A (en) 1985-07-08 1988-05-10 Trw Inc. High-extraction efficiency annular resonator
NO156916C (no) 1985-07-10 1987-12-16 Harald B Steen Stroemningskammer for vaeskestroemsfotometer.
NO156917C (no) 1985-07-16 1987-12-16 Harald B Steen Anordning for maaling av biologiske cellers lysspredning i vaeskestroemsfotometere.
US4989977A (en) 1985-07-29 1991-02-05 Becton, Dickinson And Company Flow cytometry apparatus with improved light beam adjustment
US4794086A (en) 1985-11-25 1988-12-27 Liquid Air Corporation Method for measurement of impurities in liquids
US4770992A (en) 1985-11-27 1988-09-13 Den Engh Gerrit J Van Detection of specific DNA sequences by flow cytometry
US4999283A (en) 1986-01-10 1991-03-12 University Of Kentucky Research Foundation Method for x and y spermatozoa separation
US5447841A (en) 1986-01-16 1995-09-05 The Regents Of The Univ. Of California Methods for chromosome-specific staining
US5756696A (en) 1986-01-16 1998-05-26 Regents Of The University Of California Compositions for chromosome-specific staining
US4710635A (en) 1986-04-14 1987-12-01 Becton, Dickinson And Company Dual laser excitation from single laser source
NL8601000A (nl) 1986-04-21 1987-11-16 Jan Greve T H Twente Afdeling Het gebruik van gepolariseerd licht in stromingscytometrie.
JPS62274238A (ja) 1986-05-22 1987-11-28 ベクトン・デイツキンソン・アンド・カンパニ− フロ−サイトメトリ−装置に用いる光学的結合用ゲル
US4790653A (en) 1986-05-22 1988-12-13 Becton Dickinson And Company Housing for a flow cytometry apparatus with particle unclogging feature
US4786165A (en) 1986-07-10 1988-11-22 Toa Medical Electronics Co., Ltd. Flow cytometry and apparatus therefor
US4867908A (en) 1986-08-29 1989-09-19 Becton, Dickinson And Company Method and materials for calibrating flow cytometers and other analysis instruments
US4704891A (en) 1986-08-29 1987-11-10 Becton, Dickinson And Company Method and materials for calibrating flow cytometers and other analysis instruments
FR2609885B1 (fr) 1987-01-22 1989-04-14 Cassou Robert Instrument pour l'insemination artificielle, le transfert d'embryons ou le prelevement de liquides folliculaires chez les mammiferes
US4780451B1 (en) 1987-01-23 1995-04-04 Asua International Inc Composition and method for producing superovulation in cattle
US5162306A (en) 1987-01-23 1992-11-10 Donaldson Lloyd E Composition and method for producing superovulation in mammals
EP0279000B1 (en) 1987-02-17 1993-07-21 Ratcom, Inc. Flow cytometry
US4764013A (en) 1987-03-23 1988-08-16 The United States Of America As Represented By The United States Department Of Energy Interferometric apparatus and method for detection and characterization of particles using light scattered therefrom
US4765737A (en) 1987-03-30 1988-08-23 Cornell Research Foundation Cell size measurements using light in flow cytometry and cell sorting
US5021244A (en) 1988-12-06 1991-06-04 Cytogam, Inc. Sex-associated membrane antibodies and their use for increasing the probability that offspring will be of a desired sex
EP0286088B1 (en) 1987-04-08 1994-09-14 Hitachi, Ltd. A sheath flow type flow-cell device
US5346990A (en) 1987-04-08 1994-09-13 Cytogam, Inc. Sex-associated membrane proteins and methods for increasing the probability that offspring will be of a desired sex
JPS63262565A (ja) 1987-04-20 1988-10-28 Hitachi Ltd フロ−セル
DE3851176T3 (de) 1987-04-27 1999-02-11 Laser Sensor Technology Inc Vorrichtung und Verfahren zur Untersuchung von Teilchen.
EP0289677A3 (en) 1987-04-27 1989-05-10 Fritz K. Preikschat Apparatus and method for particle analysis
FR2614626B1 (fr) 1987-04-30 1989-07-21 Ranoux Claude Conteneur pour fecondation des ovocytes et replacement des embryons chez l'homme et l'animal
JP2642632B2 (ja) 1987-07-03 1997-08-20 株式会社日立製作所 微粒子計測装置および微粒子計測方法
GB8716285D0 (en) 1987-07-10 1987-08-19 Medical Res Council Light collecting device
US4979093A (en) 1987-07-16 1990-12-18 Cavro Scientific Instruments XYZ positioner
US4987539A (en) 1987-08-05 1991-01-22 Stanford University Apparatus and method for multidimensional characterization of objects in real time
US4796788A (en) 1987-08-26 1989-01-10 Liqui-Box Corporation Bag-in-box packaging and dispensing of substances which will not readily flow by gravity
US4758729A (en) 1987-08-28 1988-07-19 Spectra-Physics, Inc. Apparatus and method for measuring the included angle of a reflective cone
DE3832901A1 (de) 1987-10-02 1989-04-20 Hitachi Ltd Teilchenmessvorrichtung
US4793705A (en) 1987-10-07 1988-12-27 The United States Of America As Represented By The United States Department Of Energy Single molecule tracking
US4831385A (en) 1987-10-14 1989-05-16 Burlington Industries, Inc. Vacuum tray fluid-jet start-up system
US4980277A (en) 1987-10-16 1990-12-25 Cultor Ltd. Cryoprotectant solution and method
US5712807A (en) 1987-10-21 1998-01-27 Bangham; James Andrew Pulse analyzing method and apparatus
US5789155A (en) 1987-10-30 1998-08-04 California Institute Of Technology Process for identifying nucleic acids and triple helices formed thereby
GB8726304D0 (en) 1987-11-10 1987-12-16 Secr Defence Particle asymmetry analyser
GB8726305D0 (en) 1987-11-10 1987-12-16 Secr Defence Portable particle analysers
US4845025A (en) 1987-11-10 1989-07-04 Coulter Corporation Biological sample mixing apparatus and method
AU2620488A (en) 1987-11-10 1989-06-01 Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland, The Particle monitoring system
US5040890A (en) 1987-11-25 1991-08-20 Becton, Dickinson And Company Sheathed particle flow controlled by differential pressure
US4887721A (en) 1987-11-30 1989-12-19 The United States Of America As Represented By The United States Department Of Energy Laser particle sorter
US4988619A (en) 1987-11-30 1991-01-29 United States Department Of Energy Flow cytometry apparatus
US4936465A (en) 1987-12-07 1990-06-26 Zoeld Tibor Method and apparatus for fast, reliable, and environmentally safe dispensing of fluids, gases and individual particles of a suspension through pressure control at well defined parts of a closed flow-through system
US5219729A (en) 1988-02-25 1993-06-15 Serono Laboratories, Inc. Fertility assay
US5622820A (en) 1988-03-10 1997-04-22 City Of Hope Method for amplification and detection of RNA and DNA sequences
US4836038A (en) 1988-03-18 1989-06-06 Aim Instruments Ltd. Automated sampler-injector apparatus and method for sampling a quantity of sample and testing portions of said quantity
US5057413A (en) 1988-06-13 1991-10-15 Becton, Dickinson And Company Method for discriminating between intact and damaged cells in a sample
US5070080A (en) 1988-08-10 1991-12-03 Fahim Mostafa S Method of inhibiting generation, maturation, motility and viability of sperm with minerals in bioavailable form
JPH0718785B2 (ja) 1988-09-19 1995-03-06 株式会社日立製作所 フローセル装置
JP2635126B2 (ja) 1988-09-30 1997-07-30 東亜医用電子株式会社 核の分葉指数を求めるための粒子分析装置及び方法
JP2635125B2 (ja) 1988-09-30 1997-07-30 東亜医用電子株式会社 核の分葉指数を求めるための粒子分析装置及び方法
JPH02168160A (ja) 1988-12-22 1990-06-28 Omron Tateisi Electron Co 細胞選別装置
US5726009A (en) 1989-03-20 1998-03-10 Anticancer, Inc. Native-state method and system for determining viability and proliferative capacity of tissues in vitro
JPH02289808A (ja) 1989-04-28 1990-11-29 Olympus Optical Co Ltd 照明光学系
US4981580A (en) 1989-05-01 1991-01-01 Coulter Corporation Coincidence arbitration in a flow cytomery sorting system
WO1990013303A1 (en) 1989-05-10 1990-11-15 The United States Of America, Represented By The Secretary, United States Department Of Commerce Method to preselect the sex of offspring
JP2992298B2 (ja) 1989-05-12 1999-12-20 サイトガム，インコーポレイテッド 性関連膜タンパク質および子が所望の性を有する確率を増大させるための方法
US4942305A (en) 1989-05-12 1990-07-17 Pacific Scientific Company Integrating sphere aerosol particle detector
US5055393A (en) 1989-06-13 1991-10-08 Salk Institute Biotechnology/Industrial Associates, Inc. Prenatal sex determination of bovine cells using male-specific oligonucleotides
US4954715A (en) 1989-06-26 1990-09-04 Zoeld Tibor Method and apparatus for an optimized multiparameter flow-through particle and cell analyzer
JPH0353164A (ja) 1989-07-20 1991-03-07 Canon Inc サンプル供給装置及びこれを用いたサンプル測定装置
US5098657A (en) 1989-08-07 1992-03-24 Tsi Incorporated Apparatus for measuring impurity concentrations in a liquid
US5030002A (en) 1989-08-11 1991-07-09 Becton, Dickinson And Company Method and apparatus for sorting particles with a moving catcher tube
US5005981A (en) 1989-09-08 1991-04-09 Becton, Dickinson And Company Apparatus for method for causing vortices in a test tube
US5215376A (en) 1989-09-08 1993-06-01 Becton, Dickinson And Company Method for causing vortices in a test tube
DE69014507T2 (de) 1989-09-13 1995-04-13 Tiyoda Seisakusho Koushoku Kk Gerät zur Zellenvorbehandlung für Durchfluss-Zytometrie.
JP2808321B2 (ja) 1989-09-19 1998-10-08 東亜医用電子株式会社 細胞分析方法及び装置
US5072382A (en) 1989-10-02 1991-12-10 Kamentsky Louis A Methods and apparatus for measuring multiple optical properties of biological specimens
US5275787A (en) 1989-10-04 1994-01-04 Canon Kabushiki Kaisha Apparatus for separating or measuring particles to be examined in a sample fluid
DE69025256T2 (de) 1989-10-11 1996-06-27 Canon Kk Gerät und Verfahren zur Trennung von Teilchen aus flüssigkeitssuspendierten Teilchen in Zusammenhang mit deren Eigenschaften
JPH03140840A (ja) 1989-10-26 1991-06-14 Hitachi Ltd 流動細胞分析装置
FR2653885B1 (fr) 1989-10-27 1994-01-14 Abx Appareil pour le comptage et la determination d'au moins une sous-population leucocytaire.
US5034613A (en) 1989-11-14 1991-07-23 Cornell Research Foundation, Inc. Two-photon laser microscopy
US5101978A (en) 1989-11-27 1992-04-07 The United States Of America As Represented By The Secretary Of The Army Fluidic sorting device for two or more materials suspended in a fluid
AU647741B2 (en) 1989-12-01 1994-03-31 Regents Of The University Of California, The Methods and compositions for chromosome-specific staining
US5274240A (en) 1990-01-12 1993-12-28 The Regents Of The University Of California Capillary array confocal fluorescence scanner and method
DE69118429T2 (de) 1990-01-26 1996-09-12 Canon Kk Verfahren zur Messung einer Spezies unter Verwendung von Fluoreszenzlicht
JP3049254B2 (ja) 1990-02-08 2000-06-05 シスメックス株式会社 ２種類の光源を備えた光学式粒子分析装置
IL93634A0 (en) 1990-03-05 1990-12-23 Galai Lab Ltd Particle size analyzer
US5153117A (en) 1990-03-27 1992-10-06 Genetype A.G. Fetal cell recovery method
US5492534A (en) 1990-04-02 1996-02-20 Pharmetrix Corporation Controlled release portable pump
US5150313A (en) 1990-04-12 1992-09-22 Regents Of The University Of California Parallel pulse processing and data acquisition for high speed, low error flow cytometry
US5076472A (en) 1990-06-13 1991-12-31 Becton, Dickinson And Company Cleaning cycle for flow cytometers
US5087295A (en) 1990-06-13 1992-02-11 Becton Dickinson And Company Cleaning cycle for flow cytometers
JPH0716392B2 (ja) 1990-06-20 1995-03-01 株式会社ピーシーシーテクノロジー 細胞融合及び融合細胞選別システム
JPH0710226B2 (ja) 1990-06-20 1995-02-08 株式会社ピーシーシーテクノロジー プロトプラスト攪拌装置並びにそれを含む細胞融合装置及びフローサイトメーター装置
US5160974A (en) 1990-06-25 1992-11-03 Flow Science, Inc. Closed sample cell for use in flow cytometry
JPH04126065A (ja) 1990-06-25 1992-04-27 Saburo Okonogi 食品素材の棒状体成型装置
US5559032A (en) 1990-06-29 1996-09-24 Pomeroy; Patrick C. Method and apparatus for post-transfer assaying of material on solid support
CA2086330A1 (en) 1990-07-09 1992-01-10 Richard C. Fry Enhanced implantation, development and maintenance of embryos using leukaemia inhibitory factor
JP2939647B2 (ja) 1990-07-24 1999-08-25 シスメックス株式会社 フローイメージングサイトメータにおける自動焦点調整方法
IE76732B1 (en) 1990-08-07 1997-11-05 Becton Dickinson Co One step test for absolute counts
US5259593A (en) 1990-08-30 1993-11-09 University Of Southern California Apparatus for droplet stream manufacturing
JPH04115136A (ja) 1990-09-05 1992-04-16 Hitachi Ltd 粒子計測装置
JPH04126080A (ja) 1990-09-14 1992-04-27 Juzo Udaka 耐熱性キシロースイソメラーゼの製造方法
JPH04126064A (ja) 1990-09-14 1992-04-27 Nitto Shokai:Kk 二重包餡食品の製造法
US5132548A (en) 1990-09-14 1992-07-21 High Yield Technology High sensitivity, large detection area particle sensor for vacuum applications
JP2957246B2 (ja) 1990-09-14 1999-10-04 大和化成株式会社 微生物起源カルボキシペプチダーゼｂ様酵素
US5204884A (en) 1991-03-18 1993-04-20 University Of Rochester System for high-speed measurement and sorting of particles
US5273527A (en) 1992-05-12 1993-12-28 Ovamed Corporation Delivery catheter
US5663048A (en) 1990-10-04 1997-09-02 University Of Calgary Y-chromosome specific polynucleotide probes for prenatal sexing
US5840482A (en) 1990-10-10 1998-11-24 The Regents Of The University Of California Y chromosome specific nucleic acid probe and method for determining the Y chromosome in situ
WO1992008120A1 (en) 1990-10-29 1992-05-14 Macquarie University Pulsed laser flow cytometry
US5116125A (en) 1990-10-31 1992-05-26 Biophos Medical Ab Fertility analyzer
DE69013366T2 (de) 1990-10-31 1995-04-20 Biophos Medical Ab Fruchtbarkeitsanalysegerät.
JP2874746B2 (ja) 1990-11-22 1999-03-24 シスメックス株式会社 フローイメージングサイトメータにおけるフローセル機構
US5991028A (en) 1991-02-22 1999-11-23 Applied Spectral Imaging Ltd. Spectral bio-imaging methods for cell classification
JPH0734012B2 (ja) 1991-02-27 1995-04-12 東亜医用電子株式会社 フローイメージサイトメータ
JP3121849B2 (ja) 1991-02-27 2001-01-09 シスメックス株式会社 フローイメージサイトメータ
US5144224A (en) 1991-04-01 1992-09-01 Larsen Lawrence E Millimeter wave flow cytometer
US5199576A (en) 1991-04-05 1993-04-06 University Of Rochester System for flexibly sorting particles
EP0515211A3 (en) 1991-05-23 1993-04-07 Becton Dickinson And Company Apparatus and method for phase resolved fluorescence lifetimes of independent and varying amplitude pulses
DE9107792U1 (zh) 1991-06-25 1991-09-12 Labotect-Labor-Technik, Goettingen, Gmbh, 3406 Bovenden, De
FR2678506B1 (fr) 1991-07-01 2000-03-10 Claude Ranoux Procede de fecondation en cycle spontane.
US5548661A (en) 1991-07-12 1996-08-20 Price; Jeffrey H. Operator independent image cytometer
US5412466A (en) 1991-07-26 1995-05-02 Toa Medical Electronics Co., Ltd. Apparatus for forming flattened sample flow for analyzing particles
DE69230902D1 (de) 1991-08-30 2000-05-18 Omron Tateisi Electronics Co Vorrichtung zur Zellenanalyse
US5488469A (en) 1991-08-30 1996-01-30 Omron Corporation Cell analyzing apparatus
US5548395A (en) 1991-09-20 1996-08-20 Toa Medical Electronics Co., Ltd. Particle analyzer
US5578449A (en) 1991-10-03 1996-11-26 Hilding Ohlsson, S.A. Procedure for the sex determination of embryos in mammals especially applied to bovine embryos
JP3212647B2 (ja) 1991-10-24 2001-09-25 シスメックス株式会社 イメージングフローサイトメータ
US5919621A (en) 1991-10-24 1999-07-06 Brown; David B. Methods for diagnosing human male infertility
IT1253226B (it) 1991-10-24 1995-07-11 Piero Serra Catetere per l'introduzione o l'aspirazione di liquidi di diversa natura in animali particolarmente per trattamenti ginecologici in bovini, equini e simili
US5866344A (en) 1991-11-15 1999-02-02 Board Of Regents, The University Of Texas System Antibody selection methods using cell surface expressed libraries
US5370842A (en) 1991-11-29 1994-12-06 Canon Kabushiki Kaisha Sample measuring device and sample measuring system
JP2593021B2 (ja) 1991-12-13 1997-03-19 伊藤ハム株式会社 ウシ胚の性の識別方法
JP3130628B2 (ja) 1992-01-30 2001-01-31 シスメックス株式会社 粒子判定装置
US5400179A (en) 1992-02-18 1995-03-21 Asahi Kogaku Kogyo Kabushiki Kaisha Optical multilayer thin film and beam splitter
ATE172791T1 (de) 1992-02-20 1998-11-15 Canon Kk Verfahren und messapparatur zur handhabung von teilchen
CA2130343C (en) 1992-02-21 2003-02-11 The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland Analysis of particle characteristics
US5558998A (en) 1992-02-25 1996-09-24 The Regents Of The Univ. Of California DNA fragment sizing and sorting by laser-induced fluorescence
US6120735A (en) 1992-02-26 2000-09-19 The Ohio States University Fractional cell sorter
JPH084601B2 (ja) * 1992-04-10 1996-01-24 岡山県 精子凍結方法及び精子凍結装置
US5298967A (en) 1992-06-02 1994-03-29 Pacific Scientific Company Measurement of concentrations of dissolved solvent
JP3145486B2 (ja) 1992-06-12 2001-03-12 シスメックス株式会社 イメージングフローサイトメータ
JPH07509153A (ja) 1992-07-13 1995-10-12 ポール・コーポレーション 生物学的流体を処理するための自動化されたシステムおよび方法
JP3215175B2 (ja) 1992-08-10 2001-10-02 シスメックス株式会社 粒子分析装置
US5315122A (en) 1992-08-25 1994-05-24 Becton, Dickinson And Company Apparatus and method for fluorescent lifetime measurement
DK0662124T3 (da) 1992-09-03 2002-10-07 Univ California Højhastigheds-flowcytometrisk separation af levedygtige pattedyrceller
US5466572A (en) 1992-09-03 1995-11-14 Systemix, Inc. High speed flow cytometric separation of viable cells
US5736410A (en) 1992-09-14 1998-04-07 Sri International Up-converting reporters for biological and other assays using laser excitation techniques
US5275933A (en) 1992-09-25 1994-01-04 The Board Of Trustees Of The Leland Stanford Junior University Triple gradient process for recovering nucleated fetal cells from maternal blood
US5371585A (en) 1992-11-10 1994-12-06 Pacific Scientific Company Particle detecting instrument with sapphire detecting cell defining a rectangular flow path
US5359907A (en) 1992-11-12 1994-11-01 Horiba Instruments, Inc. Method and apparatus for dry particle analysis
US5395588A (en) 1992-12-14 1995-03-07 Becton Dickinson And Company Control of flow cytometer having vacuum fluidics
WO1994016499A2 (en) 1993-01-16 1994-07-21 Bangham James A Signal processing system
JP2525713B2 (ja) 1993-01-19 1996-08-21 農林水産省畜産試験場長 低分子チオ―ル化合物を用いた牛胚の培養法および輸送法
US5467189A (en) 1993-01-22 1995-11-14 Venturedyne, Ltd. Improved particle sensor and method for assaying a particle
JP3052665B2 (ja) 1993-01-26 2000-06-19 株式会社日立製作所 フローセル装置
CA2113957A1 (en) 1993-01-29 1994-07-30 University Of Guelph Nucleotide sequences for bovine sex determination
IL108497A0 (en) 1993-02-01 1994-05-30 Seq Ltd Methods and apparatus for dna sequencing
US5453575A (en) 1993-02-01 1995-09-26 Endosonics Corporation Apparatus and method for detecting blood flow in intravascular ultrasonic imaging
US5367474A (en) 1993-02-08 1994-11-22 Coulter Corporation Flow cytometer
US5547849A (en) 1993-02-17 1996-08-20 Biometric Imaging, Inc. Apparatus and method for volumetric capillary cytometry
US5556764A (en) 1993-02-17 1996-09-17 Biometric Imaging, Inc. Method and apparatus for cell counting and cell classification
US5563059A (en) 1993-02-23 1996-10-08 Genentech, Inc. Use of human inhibin and human activin to increase the number of mature primate oocytes
EP0648348B1 (en) 1993-03-01 1999-04-28 General Signal Corporation Variable annular illuminator for photolithographic projection imager.
NO930980L (no) 1993-03-18 1994-09-19 Flowtech As Optisk konfigurasjon for væskeströmcytofotometer
US5658751A (en) 1993-04-13 1997-08-19 Molecular Probes, Inc. Substituted unsymmetrical cyanine dyes with selected permeability
US5494795A (en) 1993-05-05 1996-02-27 The United States Of America As Represented By The Secretary Of The Navy Specific oligonucleotide primers for detection of pathogenic campylobacter bacteria by polymerase chain reaction
US5439578A (en) 1993-06-03 1995-08-08 The Governors Of The University Of Alberta Multiple capillary biochemical analyzer
DE69418248T2 (de) 1993-06-03 1999-10-14 Hamamatsu Photonics Kk Optisches Laser-Abtastsystem mit Axikon
WO1994028810A1 (en) 1993-06-04 1994-12-22 Kwahak International Co., Ltd. Artificial insemination and embryo transfer device
NO932088L (no) 1993-06-08 1995-01-05 Oddbjoern Gjelsnes Anordning for anvendelse ved væskeströmscytometri
US5483469A (en) 1993-08-02 1996-01-09 The Regents Of The University Of California Multiple sort flow cytometer
US5464581A (en) 1993-08-02 1995-11-07 The Regents Of The University Of California Flow cytometer
US6328071B1 (en) 1993-08-06 2001-12-11 Cary Austin Well pressure tank
DE69434551T2 (de) 1993-09-16 2006-06-14 Sysmex Corp Teilchen-Analysegerät
US5503994A (en) 1993-10-08 1996-04-02 The Board Of Trustees Of The Leland Stanford Junior University System for sample detection with compensation for difference in sensitivity to detection of components moving at different velocities
US5480774A (en) 1993-10-14 1996-01-02 A/F Protein, Inc. Determination of genomic sex in salmonids
JP3290786B2 (ja) 1993-11-26 2002-06-10 シスメックス株式会社 粒子分析装置
GB9324938D0 (en) 1993-12-04 1994-01-26 Atomic Energy Authority Uk Aerosol generator
FI96452C (fi) 1994-01-26 1996-06-25 Pekka Haenninen Menetelmä väriaineiden virittämiseksi
DE69430587T2 (de) 1994-02-28 2003-01-09 Sunkyong Ind Ltd Pyrimidin-acyclonukleosid-derivate
EP0708177B1 (en) 1994-03-25 2005-09-28 Marukin Chuyu Co., Ltd. Epimerase
US5475487A (en) 1994-04-20 1995-12-12 The Regents Of The University Of California Aqueous carrier waveguide in a flow cytometer
DE4414940C2 (de) 1994-04-28 1998-07-02 Pekka Haenninen Lumineszenz-Rastermikroskop mit zwei Photonen Anregung
US5595866A (en) 1994-05-27 1997-01-21 Methodist Hospital Of Indiana, Inc. Step-wise method to remove cryoprotectant from sperm
DE4419894A1 (de) 1994-06-07 1995-12-14 Gip Medizin Technik Gmbh Endoskopische Punktionsnadelvorrichtung
EP0687956B2 (de) 1994-06-17 2005-11-23 Carl Zeiss SMT AG Beleuchtungseinrichtung
US5891734A (en) 1994-08-01 1999-04-06 Abbott Laboratories Method for performing automated analysis
US5601234A (en) 1994-08-01 1997-02-11 Abbott Laboratories Fluid nozzle and method of introducing a fluid
JP3375203B2 (ja) 1994-08-08 2003-02-10 シスメックス株式会社 細胞分析装置
FR2723735B1 (fr) 1994-08-18 1996-10-31 Abx Sa Boitier a branchement automatique pour distribution de reactifs dans un appareil notamment un analyseur hematologique.
US5790692A (en) 1994-09-07 1998-08-04 Jeffrey H. Price Method and means of least squares designed filters for image segmentation in scanning cytometry
US20020186874A1 (en) 1994-09-07 2002-12-12 Jeffrey H. Price Method and means for image segmentation in fluorescence scanning cytometry
US5700692A (en) 1994-09-27 1997-12-23 Becton Dickinson And Company Flow sorter with video-regulated droplet spacing
IL115327A (en) 1994-10-07 2000-08-13 Bayer Ag Diaphragm pump
US5934885A (en) 1994-10-07 1999-08-10 Bayer Corporation Reagent pump assembly
US5602349A (en) 1994-10-14 1997-02-11 The University Of Washington Sample introduction system for a flow cytometer
US5643796A (en) 1994-10-14 1997-07-01 University Of Washington System for sensing droplet formation time delay in a flow cytometer
US6861265B1 (en) 1994-10-14 2005-03-01 University Of Washington Flow cytometer droplet formation system
DE69532416T2 (de) 1994-10-14 2004-12-02 University Of Washington, Seattle Methode und vorrichtung zur tröpfchenerzeugung mit hoher rate bei der durchfluss-cytometrie
US5602039A (en) 1994-10-14 1997-02-11 The University Of Washington Flow cytometer jet monitor system
US5495719A (en) 1994-11-14 1996-03-05 Gray, Jr.; Carl O. Method of preserving spermatozoa
JP3347495B2 (ja) 1994-11-14 2002-11-20 シスメックス株式会社 粒子分析装置
US5514537A (en) 1994-11-28 1996-05-07 Board Of Supervisors Of Louisiana State University And Agricultural And Mechanical College Process and apparatus for sorting spermatozoa
DE69530072T2 (de) 1994-12-08 2004-03-04 Molecular Dynamics, Sunnyvale System zur fluoreszenzabbildung unter verwendung eines objektivs mit makroabtastung
US5632754A (en) 1994-12-23 1997-05-27 Devices For Vascular Intervention Universal catheter with interchangeable work element
EP0720012B1 (en) 1994-12-26 2004-09-08 Sysmex Corporation Flow cytometer
US5835262A (en) 1994-12-28 1998-11-10 Research Development Corporation Of Japan Multi-wavelength optical microscope
FI98765C (fi) 1995-01-16 1997-08-11 Erkki Soini Virtaussytometrinen menetelmä ja laite
US5793485A (en) 1995-03-20 1998-08-11 Sandia Corporation Resonant-cavity apparatus for cytometry or particle analysis
US5608519A (en) 1995-03-20 1997-03-04 Gourley; Paul L. Laser apparatus and method for microscopic and spectroscopic analysis and processing of biological cells
US5620842A (en) 1995-03-29 1997-04-15 Becton Dickinson And Company Determination of the number of fluorescent molecules on calibration beads for flow cytometry
US5786560A (en) 1995-03-31 1998-07-28 Panasonic Technologies, Inc. 3-dimensional micromachining with femtosecond laser pulses
US5699152A (en) * 1995-04-03 1997-12-16 Alltrista Corporation Electro-optical inspection system and method
US5528045A (en) 1995-04-06 1996-06-18 Becton Dickinson And Company Particle analyzer with spatially split wavelength filter
US5682038A (en) 1995-04-06 1997-10-28 Becton Dickinson And Company Fluorescent-particle analyzer with timing alignment for analog pulse subtraction of fluorescent pulses arising from different excitation locations
US5641457A (en) 1995-04-25 1997-06-24 Systemix Sterile flow cytometer and sorter with mechanical isolation between flow chamber and sterile enclosure
US5687727A (en) 1995-05-01 1997-11-18 Danforth Biomedical Incorporated Catheter adaptor with slitting blade and improved manual control and method of use
EP0957797A4 (en) 1995-05-09 2001-05-16 Univ Missouri SYSTEM FOR INITIATING A FLUID IN THE BOTTOM OF AN ANIMAL
FR2734637B1 (fr) 1995-05-24 1997-08-14 Abx Sa Dispositif d'inspection optique d'un fluide, notamment pour analyses hematologiques
US5798276A (en) 1995-06-07 1998-08-25 Molecular Probes, Inc. Reactive derivatives of sulforhodamine 101 with enhanced hydrolytic stability
US5650847A (en) 1995-06-14 1997-07-22 Erkki Soini Method and device for determination of parameters of individual microparticles
US5716852A (en) 1996-03-29 1998-02-10 University Of Washington Microfabricated diffusion-based chemical sensor
US6454945B1 (en) 1995-06-16 2002-09-24 University Of Washington Microfabricated devices and methods
US5589457A (en) 1995-07-03 1996-12-31 Ausa International, Inc. Process for the synchronization of ovulation
US6411835B1 (en) 1997-01-13 2002-06-25 Medispectra, Inc. Spectral volume microprobe arrays
DE69634449T2 (de) 1995-08-11 2006-01-05 University Of Guelph, Guelph Verfahren zum identifizieren von geschlechtsspezifischen und speziesspezifischen molekülen, mit verfahren identifizierte moleküle und verwendung der moleküle
JP2001520637A (ja) 1995-09-06 2001-10-30 ザ・リサーチ・ファンデーション・オブ・ステート・ユニバーシティ・オブ・ニューヨーク 二光子アップコンバーティング色素および応用
DE19533092A1 (de) 1995-09-07 1997-03-13 Basf Ag Vorrichtung zur parallelisierten Zweiphotonen-Fluoreszenz-Korrelations-Spektroskopie (TPA-FCS) und deren Verwendung zum Wirkstoff-Screening
US6329158B1 (en) 1995-09-15 2001-12-11 Becton Dickinson And Company Use of dimly fluorescing nucleic acid dyes in the identification of nucleated cells
US6040139A (en) 1995-09-19 2000-03-21 Bova; G. Steven Laser cell purification system
US5726751A (en) 1995-09-27 1998-03-10 University Of Washington Silicon microchannel optical flow cytometer
US5780230A (en) 1995-10-06 1998-07-14 Coriell Institute For Medical Research Compositions and methods for human sperm activation and quantitative assessment of human sperm genome replication
US5736330A (en) 1995-10-11 1998-04-07 Luminex Corporation Method and compositions for flow cytometric determination of DNA sequences
US6117068A (en) 1995-10-19 2000-09-12 Elite Genetics, Inc Artificial insemination system
EP0857203B1 (en) 1995-10-19 2008-08-06 Bio-Origyn LLC Methods and compositions to improve germ cell and embryo survival and function
DE69529538T2 (de) 1995-11-09 2003-11-06 Medical Res Council London Verwendung von cyb-medium zum transport und lagern von sperma
DE19549015C1 (de) 1995-12-28 1997-04-03 Siemens Ag Verfahren und Anordnung zur Überwachung eines abreißenden Flüssigkeitstrahls
JP3584108B2 (ja) 1996-01-08 2004-11-04 キヤノン株式会社 レンズ鏡筒
US6641708B1 (en) 1996-01-31 2003-11-04 Board Of Regents, The University Of Texas System Method and apparatus for fractionation using conventional dielectrophoresis and field flow fractionation
WO1997029354A1 (de) 1996-02-05 1997-08-14 Bayer Aktiengesellschaft Verfahren und vorrichtung zum sortieren und zur gewinnung von planar ausgebrachten biologischen objekten wie biologische zellen bzw. zellorganellen, histologischen schnitten, chromosomenteilchen etc. mit laserstrahlen
BR9600722A (pt) 1996-02-14 1997-12-30 Jorge Antonio Rodrigues Claro Bomba de infusão para contidos em bolsas plásticas flexíveis
WO1997030338A1 (en) 1996-02-16 1997-08-21 Inphocyte, Inc. System and method for rapid analysis of cells using spectral cytometry
JP3127111B2 (ja) 1996-02-22 2001-01-22 株式会社日立製作所 フロー式粒子画像解析方法および装置
US6090947A (en) * 1996-02-26 2000-07-18 California Institute Of Technology Method for the synthesis of pyrrole and imidazole carboxamides on a solid support
US6143901A (en) * 1996-07-31 2000-11-07 Genesoft, Inc. Complex formation between dsDNA and pyrrole imidazole polyamides
US5701012A (en) 1996-03-19 1997-12-23 Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of National Defence Fluorescent biological particle detection system
US5895922A (en) 1996-03-19 1999-04-20 Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of National Defence Fluorescent biological particle detection system
US5747349A (en) 1996-03-20 1998-05-05 University Of Washington Fluorescent reporter beads for fluid analysis
JP3590470B2 (ja) 1996-03-27 2004-11-17 アルプス電気株式会社 洗浄水生成方法および洗浄方法ならびに洗浄水生成装置および洗浄装置
JP3640461B2 (ja) 1996-04-03 2005-04-20 シスメックス株式会社 粒子分析装置
US5707808A (en) 1996-04-15 1998-01-13 The Regents Of The University Of California Optical selection and collection of DNA fragments
DE69737513T2 (de) 1996-04-25 2007-12-13 Genicon Sciences Corp., San Diego Teilchenförmiges markierungsmittel verwendendes analytassay
WO1997043620A1 (en) 1996-05-15 1997-11-20 International Remote Imaging Systems, Inc. Selectively emphasizing particles of interest from a fluid sample for analysis
JP2963393B2 (ja) 1996-06-14 1999-10-18 浜松ホトニクス株式会社 光電子増倍管用電圧分割回路
US5846737A (en) 1996-07-26 1998-12-08 Molecular Probes, Inc. Conjugates of sulforhodamine fluorophores with enhanced fluorescence
US5909278A (en) 1996-07-29 1999-06-01 The Regents Of The University Of California Time-resolved fluorescence decay measurements for flowing particles
EP0822401A3 (en) 1996-07-30 1999-05-06 Bayer Corporation Hydraulic system for a hematology analytical instrument
US5844685A (en) 1996-07-30 1998-12-01 Bayer Corporation Reference laser beam sampling apparatus
US5872627A (en) 1996-07-30 1999-02-16 Bayer Corporation Method and apparatus for detecting scattered light in an analytical instrument
US5883378A (en) 1996-07-30 1999-03-16 Bayer Corporation Apparatus and methods for transmitting electrical signals indicative of optical interactions between a light beam and a flowing suspension of particles
US6042249A (en) 1996-07-30 2000-03-28 Bayer Corporation Illuminator optical assembly for an analytical instrument and methods of alignment and manufacture
US6074827A (en) 1996-07-30 2000-06-13 Aclara Biosciences, Inc. Microfluidic method for nucleic acid purification and processing
US5719667A (en) 1996-07-30 1998-02-17 Bayer Corporation Apparatus for filtering a laser beam in an analytical instrument
DE69739456D1 (de) 1996-07-30 2009-07-30 Siemens Healthcare Diagnostics Optisches System für Blutanalysegerät
US5745308A (en) 1996-07-30 1998-04-28 Bayer Corporation Methods and apparatus for an optical illuminator assembly and its alignment
US5998140A (en) * 1996-07-31 1999-12-07 The Scripps Research Institute Complex formation between dsDNA and oligomer of cyclic heterocycles
US5804436A (en) 1996-08-02 1998-09-08 Axiom Biotechnologies, Inc. Apparatus and method for real-time measurement of cellular response
EP1012450B1 (en) 1996-08-07 2004-10-20 Cuno Incorporated Additive dispensing apparatus
AU4113297A (en) 1996-09-04 1998-03-26 Technical University Of Denmark A micro flow system for particle separation and analysis
ATE267596T1 (de) 1996-09-06 2004-06-15 Medarex Inc Cyanidinzusammensetzungen und deren therapeutische und diagnostische verwendungen
US5873254A (en) 1996-09-06 1999-02-23 Interface Multigrad Technology Device and methods for multigradient directional cooling and warming of biological samples
US6221654B1 (en) 1996-09-25 2001-04-24 California Institute Of Technology Method and apparatus for analysis and sorting of polynucleotides based on size
US5759767A (en) 1996-10-11 1998-06-02 Joseph R. Lakowicz Two-photon and multi-photon measurement of analytes in animal and human tissues and fluids
US6002471A (en) 1996-11-04 1999-12-14 California Institute Of Technology High resolution scanning raman microscope
US5799830A (en) 1996-11-08 1998-09-01 Carroll; David C. Pressure vessel access port
US5696157A (en) 1996-11-15 1997-12-09 Molecular Probes, Inc. Sulfonated derivatives of 7-aminocoumarin
CA2279574C (en) 1997-01-31 2007-07-24 The Horticulture & Food Research Institute Of New Zealand Ltd. Optical apparatus
US6753161B2 (en) 1997-03-27 2004-06-22 Oncosis Llc Optoinjection methods
US6534308B1 (en) 1997-03-27 2003-03-18 Oncosis, Llc Method and apparatus for selectively targeting specific cells within a mixed cell population
US5874266A (en) 1997-03-27 1999-02-23 Palsson; Bernhard O. Targeted system for removing tumor cells from cell populations
GB9707096D0 (en) 1997-04-08 1997-05-28 Smithkline Beecham Plc Novel device
JP2968231B2 (ja) 1997-04-11 1999-10-25 株式会社荏原製作所 空調システム
US6133995A (en) 1997-05-09 2000-10-17 Sysmex Corporation Particle measuring apparatus
US6050935A (en) 1997-05-09 2000-04-18 Biofertec Container assembly for intravaginal fertilization and culture and embryo transfer and method of intravaginal fertilization and culture employing such a container
EP0988523B1 (en) 1997-06-09 2013-08-14 EMD Millipore Corporation Method and apparatus for detecting microparticles in fluid samples
WO1998059233A1 (en) 1997-06-23 1998-12-30 Luminex Corporation Interlaced lasers for multiple fluorescence measurement
DK0894498T3 (da) 1997-07-01 2006-08-28 Vlp Watertown Ltd Partnership Fremgangsmåde til bestemmelse af kön af pattedyr-afkom
US6111398A (en) 1997-07-03 2000-08-29 Coulter International Corp. Method and apparatus for sensing and characterizing particles
BR9704313A (pt) 1997-07-08 1999-04-06 Alves Elias Walter Utilização de ig-y de ovo de galinha associado a anticorpos monoclonais contra antigenos sexo especifico na imunosexagem de espermatozóides de bovinos
US5899848A (en) 1997-07-14 1999-05-04 Haubrich; Mark A. Device and process for artificial insemination of animals
US5876942A (en) 1997-07-24 1999-03-02 National Science Council Of Republic Of China Process for sexing cow embryos
US5985216A (en) 1997-07-24 1999-11-16 The United States Of America, As Represented By The Secretary Of Agriculture Flow cytometry nozzle for high efficiency cell sorting
US5985538A (en) 1997-08-01 1999-11-16 Saint Barnabas Medical Center Cryopreservation and cell culture medium comprising less than 50 mM sodium ions and greater than 100 mM choline salt
US5819948A (en) 1997-08-21 1998-10-13 Van Den Engh; Gerrit J. Particle separating apparatus and method
US6003678A (en) 1997-08-21 1999-12-21 University Of Washington Particle separating apparatus and method
US5880474A (en) 1997-08-29 1999-03-09 Becton Dickinson And Company Multi-illumination-source flow particle analyzer with inter-location emissions crosstalk cancelation
AU9247598A (en) 1997-09-22 1999-04-12 University Of Guelph Reduction of sperm sensitivity to chilling
US6540895B1 (en) 1997-09-23 2003-04-01 California Institute Of Technology Microfabricated cell sorter for chemical and biological materials
US6322901B1 (en) * 1997-11-13 2001-11-27 Massachusetts Institute Of Technology Highly luminescent color-selective nano-crystalline materials
CA2310672A1 (en) 1997-11-19 1999-05-27 University Of Washington High throughput optical scanner
US6086574A (en) 1997-11-21 2000-07-11 Hyclone Laboratories, Inc. Fluid delivery systems with diptube connector
US5895764A (en) 1997-11-24 1999-04-20 University Of New Mexico Controlled sheath flow injection cytometry
US6207392B1 (en) 1997-11-25 2001-03-27 The Regents Of The University Of California Semiconductor nanocrystal probes for biological applications and process for making and using such probes
US5990479A (en) 1997-11-25 1999-11-23 Regents Of The University Of California Organo Luminescent semiconductor nanocrystal probes for biological applications and process for making and using such probes
US6221671B1 (en) 1997-12-12 2001-04-24 Chemunex S.A. Digital flow cytometer and method
US6149867A (en) 1997-12-31 2000-11-21 Xy, Inc. Sheath fluids and collection systems for sex-specific cytometer sorting of sperm
US6071689A (en) 1997-12-31 2000-06-06 Xy, Inc. System for improving yield of sexed embryos in mammals
US6617107B1 (en) 1998-02-03 2003-09-09 Xy, Inc. Specific oligonucleotide primers for detection of bovine male chromosome presence by polymerase chain reaction and method
KR20010041113A (ko) 1998-02-20 2001-05-15 헤릭호프, 리사 분류용 플로우 사이토미터의 진동시스템
US6746873B1 (en) 1998-02-20 2004-06-08 Xy, Inc. Vibratory system for a sorting flow cytometer
US6154276A (en) 1998-02-23 2000-11-28 The Regents Of The University Of California Waveguide detection of right-angle-scattered light in flow cytometry
US6211477B1 (en) 1998-02-26 2001-04-03 Becton Dickinson And Company Electrostatic deceleration system for flow cytometer
US6248590B1 (en) 1998-02-27 2001-06-19 Cytomation, Inc. Method and apparatus for flow cytometry
US6042025A (en) * 1998-03-13 2000-03-28 Smith Et Al. Two hole dispenser with baffles
DE69912935T2 (de) 1998-03-16 2004-09-02 Partec Partikelzählgeräte GmbH Elektronisches gerät zur präzisen abgabe kleiner flüssigkeitsmengen
JP3594794B2 (ja) 1998-03-24 2004-12-02 独立行政法人 科学技術振興機構 ナノ秒時間ゲート分光診断装置
US6642018B1 (en) * 1998-03-27 2003-11-04 Oncosis Llc Method for inducing a response in one or more targeted cells
EP1068509A4 (en) 1998-03-30 2005-05-25 Bioshaf FLOW CYTOMETER FOR THE ANALYSIS OF GENERAL DIAGNOSTIC FACTORS IN CELLS AND BIOLOGICAL FLUIDS
US6175409B1 (en) 1999-04-02 2001-01-16 Symyx Technologies, Inc. Flow-injection analysis and variable-flow light-scattering methods and apparatus for characterizing polymers
JP3350442B2 (ja) 1998-04-09 2002-11-25 科学技術振興事業団 顕微鏡システム
CA2328408C (en) 1998-05-14 2005-02-15 Luminex Corporation Zero dead time architecture and method for flow cytometer
WO1999060397A1 (en) 1998-05-18 1999-11-25 University Of Washington Liquid analysis cartridge
JP2002528699A (ja) 1998-05-22 2002-09-03 カリフォルニア インスティチュート オブ テクノロジー 微細製作細胞分類器
US6079836A (en) 1998-07-20 2000-06-27 Coulter International Corp. Flow cytometer droplet break-off location adjustment mechanism
NZ509434A (en) * 1998-07-30 2004-03-26 Univ Colorado State Res Found Equine system for non-surgical artificial insemination
US20040107150A1 (en) 1998-07-31 2004-06-03 Chata Biosystems, Inc. Of Colorado Apparatus and method with vessel for containing/transporting a fluent substance
US6729369B2 (en) 1998-07-31 2004-05-04 Chata Biosystems, Inc. Vessel for containing/transporting a fluent substance
DE29813921U1 (de) 1998-08-04 1998-10-08 Kisfeld Alfons Vorrichtung zur Besamung von Tieren, insbesondere Sauen
US6309815B1 (en) * 1998-08-07 2001-10-30 University Of Kansas Medical Center Composition and method for preparation, storage and activation of large populations of immotile sperm
WO2000009113A1 (en) 1998-08-17 2000-02-24 Trout William E Use of zeranol to modulate reproductive cycles
AU754644B2 (en) 1998-08-21 2002-11-21 Union Biometrica, Inc. Instrument for selecting and depositing multicellular organisms and other large objects
US6326144B1 (en) 1998-09-18 2001-12-04 Massachusetts Institute Of Technology Biological applications of quantum dots
US6495333B1 (en) 1998-09-22 2002-12-17 Becton Dickinson And Company Flow cytometric, whole blood dendritic cell immune function assay
US6208411B1 (en) 1998-09-28 2001-03-27 Kla-Tencor Corporation Massively parallel inspection and imaging system
US6707555B1 (en) 1998-10-15 2004-03-16 Sysmex Corporation Optical information measuring apparatus
US6423505B1 (en) 1998-12-03 2002-07-23 Becton Dickinson And Company Methods and reagents for quantitation of HLA-DR and CD11b expression on peripheral blood cells
US7116407B2 (en) 1998-12-15 2006-10-03 Union Biometrica, Inc. System for axial pattern analysis of multicellular organisms
US6128133A (en) 1998-12-22 2000-10-03 Lucent Technologies Inc. Optical beamsplitter
JP2000206032A (ja) 1999-01-06 2000-07-28 Bayer Corp 可変割合体積粒子計数装置及び試料の特性を定める方法
US6256096B1 (en) 1999-01-11 2001-07-03 Softray Flow cytometry apparatus and method
US20010006416A1 (en) 1999-01-11 2001-07-05 Johnson Paul E. Ribbon flow cytometry apparatus and methods
US6150119A (en) 1999-01-19 2000-11-21 Caliper Technologies Corp. Optimized high-throughput analytical system
US6580504B1 (en) 1999-01-25 2003-06-17 Amnis Corporation Multipass cavity for illumination and excitation of moving objects
US6473176B2 (en) 1999-01-25 2002-10-29 Amnis Corporation Imaging and analyzing parameters of small moving objects such as cells
US6671044B2 (en) 1999-01-25 2003-12-30 Amnis Corporation Imaging and analyzing parameters of small moving objects such as cells in broad flat flow
US6119465A (en) 1999-02-10 2000-09-19 Mullens; Patrick L. Shipping container for storing materials at cryogenic temperatures
FR2789778B1 (fr) 1999-02-12 2001-09-14 France Telecom Procede pour associer des references d'acheminement a des paquets de donnees au moyen d'une memoire trie, et routeur de paquets appliquant ce procede
JP2000239101A (ja) * 1999-02-18 2000-09-05 Japan Science & Technology Corp ラット精子の凍結保存法
US6323632B1 (en) 1999-08-13 2001-11-27 Coulter International Corp. Solid state RF oscillator-detector for flow cytometer
CH690557A5 (de) 1999-03-01 2000-10-13 Arson Ag Beleuchtungseinrichtung.
US6097485A (en) 1999-03-08 2000-08-01 Integrated Waveguides, Inc. Microchip optical transport technology for use in a personal flow cytometer
FR2791645B1 (fr) 1999-04-02 2001-06-15 Valois Sa Echantillon de produit fluide destine a la presse
US6193647B1 (en) 1999-04-08 2001-02-27 The Board Of Trustees Of The University Of Illinois Microfluidic embryo and/or oocyte handling device and method
US6793387B1 (en) 1999-05-08 2004-09-21 Chata Biosystems, Inc. Apparatus for automatic preparation of a mixture and method
FR2793495B1 (fr) 1999-05-14 2001-08-10 Imv Technologies Dilueur pour la cryoconservation de spermatozoides de bovins
FR2793708B1 (fr) 1999-05-21 2001-08-03 Valois Sa Dispositif de distribution de produit fluide
US6372506B1 (en) 1999-07-02 2002-04-16 Becton, Dickinson And Company Apparatus and method for verifying drop delay in a flow cytometer
JP3361778B2 (ja) * 1999-07-09 2003-01-07 社団法人家畜改良事業団 家畜の人工授精用精子または受精卵移植用卵子の注入器及びその操作方法
IT1307787B1 (it) 1999-07-26 2001-11-19 Univ Firenze Processo per incrementare la motilita' degli spermatozoi e spermatozoia motilita' superiore cosi' ottenuti.
DE19935766A1 (de) 1999-07-29 2001-02-01 Friedrich Schiller Uni Jena Bu Verfahren zur optischen Anregung von Fluorophor-markierter DNA und RNA
US6664550B2 (en) 1999-08-30 2003-12-16 Sandia National Laboratories Apparatus to collect, classify, concentrate, and characterize gas-borne particles
US6495366B1 (en) 1999-09-03 2002-12-17 Therakos, Inc. Uninterrupted flow pump apparatus and method
US6813017B1 (en) 1999-10-20 2004-11-02 Becton, Dickinson And Company Apparatus and method employing incoherent light emitting semiconductor devices as particle detection light sources in a flow cytometer
EP1227898A4 (en) 1999-10-21 2011-11-16 Beckman Coulter Inc SYSTEM FOR ANALYSIS BY FLOW CYTOMETRY WITH TEMPORARY DYNAMICS
US6472153B1 (en) 1999-10-26 2002-10-29 Epoch Biosciences, Inc. Hybridization-triggered fluorescent detection of nucleic acids
US7208265B1 (en) 1999-11-24 2007-04-24 Xy, Inc. Method of cryopreserving selected sperm cells
JP5087192B2 (ja) 1999-11-30 2012-11-28 インテレクソン コーポレイション 細胞群内の特定細胞に選択的に照準付けする方法及び装置
US6263745B1 (en) 1999-12-03 2001-07-24 Xy, Inc. Flow cytometer nozzle and flow cytometer sample handling methods
US6558911B1 (en) 1999-12-10 2003-05-06 Oregon Health Sciences University Sperm quality assay
DE60014440T2 (de) 2000-01-03 2005-11-17 Ibérica de Reproducción Asistida, S.L. Vorrichtung zur künstlichen befruchtung von schweinen
ES2174778T3 (es) 2000-01-14 2002-11-16 Exelixis Deutschland Gmbh Procedimiento de crioconservacion de esperma de pez cebra.
US6465169B2 (en) 2000-01-14 2002-10-15 Artemis Pharmaceuticals Gmbh Method for cryoconservation of Zebrafish sperm
IT1317724B1 (it) * 2000-01-14 2003-07-15 Istituto Sperimentale Italiano Procedimento per la produzione di embrioni non umani di sessopredeterminato ad alto valore genetico.
EP1257664A4 (en) 2000-01-28 2006-04-05 Althea Technologies Inc METHOD FOR THE ANALYSIS OF GENE EXPRESSION
US6587203B2 (en) 2000-02-17 2003-07-01 Cornell Research Foundation, Inc. Sort stream stabilizer for flow cytometer
US6618143B2 (en) 2000-02-18 2003-09-09 Idexx Laboratories, Inc. High numerical aperture flow cytometer and method of using same
US6646742B1 (en) 2000-02-19 2003-11-11 Mwi, Inc. Optical device and method for multi-angle laser light scatter
JP3587755B2 (ja) 2000-03-14 2004-11-10 シスメックス株式会社 粒子測定装置およびその方法
GB2360360B (en) 2000-03-14 2002-03-20 Univ Bristol A method of sorting cells
US6482652B2 (en) 2000-03-23 2002-11-19 The Board Of Trustees Of The Leland Stanford Junior University Biological particle sorter
MXPA02009613A (es) 2000-03-30 2004-07-30 Univ Iowa State Res Found Marcadores geneticos para caracteristicas de carne mejoradas en animales.
DK1290427T3 (en) 2000-04-11 2017-12-04 Chemometec As Method and apparatus for detecting the fluorescence of a sample
EP1147774A1 (en) 2000-04-20 2001-10-24 Stichting Dienst Landbouwkundig Onderzoek Method for improving the quality of sperm for artificial insemination of animals
WO2001085913A2 (en) 2000-05-09 2001-11-15 Xy, Inc. High purity x-chromosome bearing and y-chromosome bearing populations of spermatozoa
EP1287111A4 (en) 2000-05-19 2005-12-28 Dakocytomation Colorado Inc QUICK ENTRY SYSTEM OF MULTIMATIC SAMPLE
JP2001328901A (ja) * 2000-05-19 2001-11-27 Seatsuku Yoshitomi Kk マウス精子凍結保存用キット
US6590911B1 (en) 2000-06-02 2003-07-08 Coherent, Inc. Passively modelocked harmonic-generating laser
US6700130B2 (en) 2001-06-29 2004-03-02 Honeywell International Inc. Optical detection system for flow cytometry
US6784981B1 (en) * 2000-06-02 2004-08-31 Idexx Laboratories, Inc. Flow cytometry-based hematology system
US7351376B1 (en) 2000-06-05 2008-04-01 California Institute Of Technology Integrated active flux microfluidic devices and methods
US6503698B1 (en) 2000-06-16 2003-01-07 The United States Of America As Represented By The Secretary Of Agriculture Cryopreservation of swine embryos
BR0111618A (pt) 2000-06-12 2004-06-29 Xy Inc Sistema de controle de rebanho integrado utilizando populações isoladas de espermatozóides contendo cromosso- x e contendo coromosso y
DE10031028B4 (de) 2000-06-26 2008-09-04 Gnothis Holding Sa Verfahren zur Selektion von Partikeln
GB0016920D0 (en) 2000-07-10 2000-08-30 Univ Cambridge Tech Decondensation of DNA
AU2001286445A1 (en) 2000-08-10 2002-02-18 Gtc Biotherapeutics, Inc. Cryopreservation of sperm
US20040005582A1 (en) 2000-08-10 2004-01-08 Nanobiodynamics, Incorporated Biospecific desorption microflow systems and methods for studying biospecific interactions and their modulators
FR2813283B1 (fr) 2000-08-25 2003-02-14 Valois Sa Distributeur a pompe integree
EP1190684A1 (en) 2000-09-05 2002-03-27 Universiteit Gent Device and method for artificial insemination of bovines and other animals
US20020028434A1 (en) 2000-09-06 2002-03-07 Guava Technologies, Inc. Particle or cell analyzer and method
JP2004522418A (ja) 2000-09-08 2004-07-29 アイオワ ステート ユニバーシティ リサーチ ファウンデーション，インコーポレーティッド 新規なｐｒｋａｇ３対立遺伝子並びに生殖性及び肉質の形質の遺伝標識としてのその使用
EP2299256A3 (en) 2000-09-15 2012-10-10 California Institute Of Technology Microfabricated crossflow devices and methods
GB0023041D0 (en) 2000-09-20 2000-11-01 Univ Manchester Identification apparatus
US7208120B2 (en) 2000-09-27 2007-04-24 The Trustees Of Boston University Cellular diagnostic arrays, methods of using and processing for producing same
CN1325909C (zh) 2000-09-27 2007-07-11 清华大学 用于微粒操纵与微粒导向的装置及其使用方法
BR0005045A (pt) 2000-09-28 2002-05-14 Marcos Fernando De Resende Mat Método para sexagem de espermatozóides usando a via clássica do sistema complemento, com eliminação da via alternativa pela inativação da proteina "b"
US7258774B2 (en) 2000-10-03 2007-08-21 California Institute Of Technology Microfluidic devices and methods of use
US20040031071A1 (en) 2000-10-05 2004-02-12 Xy, Inc. System of hysteroscopic insemination of mares
AU2001236517A2 (en) 2000-10-05 2002-04-15 Xy, Inc. System of hysteroscopic insemination of mares
WO2002031467A1 (en) 2000-10-12 2002-04-18 Amnis Corporation Multipass cavity for illumination and excitation of moving objects
JP2004537712A (ja) 2000-10-18 2004-12-16 バーチャル・アレイズ・インコーポレーテッド 多重細胞分析システム
CN1328149C (zh) 2000-10-23 2007-07-25 因斯蒂尔医学技术有限公司 具有外壳和柔性内部气囊的流体分配器
AU2002220392A1 (en) 2000-11-09 2002-05-21 Universiy of Guelph Mammalian sex selection using genetic modification
SE0004777D0 (sv) 2000-12-22 2000-12-22 Amersham Pharm Biotech Ab Separation of X-and Y-sperm cells
WO2002041906A2 (en) * 2000-11-22 2002-05-30 Pharmacia Corporation Methods and apparatus for producing gender enriched sperm
US20020064809A1 (en) 2000-11-29 2002-05-30 Mutz Mitchell W. Focused acoustic ejection cell sorting system and method
CA2468774C (en) 2000-11-29 2015-06-30 George E. Seidel System for in-vitro fertilization with spermatozoa separated into x-chromosome and y-chromosome bearing populations
US6849423B2 (en) 2000-11-29 2005-02-01 Picoliter Inc Focused acoustics for detection and sorting of fluid volumes
US20020064808A1 (en) 2000-11-29 2002-05-30 Mutz Mitchell W. Focused acoustic energy for ejecting cells from a fluid
US7713687B2 (en) 2000-11-29 2010-05-11 Xy, Inc. System to separate frozen-thawed spermatozoa into x-chromosome bearing and y-chromosome bearing populations
EP1340080A2 (en) 2000-11-30 2003-09-03 The Netherlands Cancer Institute Membrane molecule indicator compositions and methods
US6576291B2 (en) 2000-12-08 2003-06-10 Massachusetts Institute Of Technology Preparation of nanocrystallites
JP2004518127A (ja) 2000-12-15 2004-06-17 サイトメーション， インコーポレイテッド 電気伝導性格納システム
US6577387B2 (en) 2000-12-29 2003-06-10 Johnson & Johnson Vision Care, Inc. Inspection of ophthalmic lenses using absorption
WO2002060880A1 (fr) 2001-01-29 2002-08-08 Universite De Geneve Derives d'acyclonucleosides pyrimidiniques, leur procede de preparation et leur utilisation
US6673095B2 (en) 2001-02-12 2004-01-06 Wound Healing Of Oklahoma, Inc. Apparatus and method for delivery of laser light
US7029916B2 (en) 2001-02-21 2006-04-18 Maxcyte, Inc. Apparatus and method for flow electroporation of biological samples
WO2002077011A2 (en) 2001-03-12 2002-10-03 The State Of Oregon Acting By And Through The State Board Of Higher Education On Behalf Of The University Of Oregon Oxidation-reduction sensitive green fluorescent protein variants
US6706163B2 (en) 2001-03-21 2004-03-16 Michael Seul On-chip analysis of particles and fractionation of particle mixtures using light-controlled electrokinetic assembly of particles near surfaces
CA2442019A1 (en) 2001-03-22 2002-10-03 Infigen, Inc. Sex-specific selection of sperm from transgenic animals
WO2002078906A2 (en) 2001-03-29 2002-10-10 Cellect Technologies Corp. Methods devices and systems for sorting and separating particles
DE60218074T2 (de) 2001-03-29 2008-02-07 Sysmex Corp. Durchflusszytometer
JP2002308897A (ja) * 2001-03-30 2002-10-23 Council Scient Ind Res 精子の運動不能性を引き起こす、インドのミミズの体腔液由来のタンパク質
WO2002082057A2 (en) 2001-04-03 2002-10-17 Micronics, Inc. Split focusing cytometer
JP2002311027A (ja) 2001-04-09 2002-10-23 Hitachi Software Eng Co Ltd ビーズ、ビーズ製造方法、フローサイトメータ及びプログラム
US6416190B1 (en) * 2001-04-27 2002-07-09 University Of Chicago Apparatus for using optical tweezers to manipulate materials
US20020186375A1 (en) 2001-05-01 2002-12-12 Asbury Charles L. Device and methods for detecting samples in a flow cytometer independent of variations in fluorescence polarization
WO2002092161A1 (en) 2001-05-10 2002-11-21 Biophan, Llc Miniaturized particle analyzer
US7345758B2 (en) 2001-05-17 2008-03-18 Cytopeia Apparatus for analyzing and sorting biological particles
EP1395374B1 (en) 2001-05-17 2013-04-17 Beckman Coulter, Inc. Flow cytometer with active automated optical alignment system
US20020182590A1 (en) 2001-05-25 2002-12-05 Vanderbilt University Determining protein function in cell culture using RNA interference
US20030048433A1 (en) 2001-06-01 2003-03-13 Jean-Marie Desjonqueres Cytometer signal processing system and method
FR2825987B1 (fr) 2001-06-19 2003-12-12 Valois Sa Distributeur de produit fluide
WO2003008937A2 (en) 2001-07-18 2003-01-30 The Regents Of The University Of Michigan Gas-focusing flow cytometer cell and flow cytometer detection system with waveguide optics
WO2003008102A1 (en) 2001-07-18 2003-01-30 The Regents Of The University Of Michigan Microfluidic gravity pump with constant flow rate
FR2828281B1 (fr) 2001-08-02 2004-12-31 Biocytex Dispositif pour l'analyse d'un echantillon notamment par cytometrie de flux
AU2002320399A1 (en) 2001-09-01 2003-03-18 Pharmacia Corporation Staining and sorting genomes and chromosomes using sequence-specific polyamides
DE10151216A1 (de) 2001-10-16 2003-04-24 Zeiss Carl Jena Gmbh Verfahren zur optischen Erfassung von charakteristischen Größen einer beleuchteten Probe
US20030078703A1 (en) 2001-10-19 2003-04-24 Surromed, Inc. Cytometry analysis system and method using database-driven network of cytometers
US6838289B2 (en) 2001-11-14 2005-01-04 Beckman Coulter, Inc. Analyte detection system
US6831279B2 (en) 2001-11-27 2004-12-14 Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of National Defence Laser diode-excited biological particle detection system
WO2003053055A1 (en) 2001-12-18 2003-06-26 Thomson Licensing S.A. Internally generated close captioning/tele-texting for set-up menus of network-capable signal processing apparatus
WO2003056330A2 (de) 2001-12-31 2003-07-10 Institut für Physikalische Hochtechnologie e.V. Zellsortiersystem zur grössenabhängigen sortierung oder separation von in einer strömenden flüssigkeit suspendierten zellen
WO2003056335A2 (de) 2001-12-31 2003-07-10 Institut für Physikalische Hochtechnologie e.V. Mikrovertiefungsarray zur grössenabhängigen sortierung oder separation von in einer strömenden flüssigkeit suspendierten zellen und verfahren zur untersuchung der funktionellen aktivität von einzelzellen
WO2003062796A1 (en) 2002-01-22 2003-07-31 Dakocytomation Denmark A/S Environmental containment system for a flow cytometer
US6698627B2 (en) 2002-02-19 2004-03-02 Valois S.A.S. Fluid dispenser
US6849394B2 (en) * 2002-02-21 2005-02-01 Minitube Of America Compositions comprising reproductive cell media and methods for using such compositions
CN1646682A (zh) 2002-02-27 2005-07-27 密执安大学评议会 从很少活动的颗粒中分选出活动颗粒的方法以及适合该方法的装置
US20030175980A1 (en) 2002-03-14 2003-09-18 Hayenga Jon W. Ribbon flow cytometry and cell sorting
US7223371B2 (en) 2002-03-14 2007-05-29 Micronics, Inc. Microfluidic channel network device
JP2004000144A (ja) 2002-03-29 2004-01-08 Aisin Seiki Co Ltd 細胞分離選別装置、細胞整列用基板
US7312085B2 (en) * 2002-04-01 2007-12-25 Fluidigm Corporation Microfluidic particle-analysis systems
CA2490334A1 (en) 2002-06-28 2004-01-08 Monsanto Technology Llc Swine genetics business system
AU2003255529B2 (en) 2002-07-10 2008-11-20 Laboratoires Serono Sa Use of compounds for increasing spermatozoa motility
EP1542529A4 (en) 2002-07-22 2006-08-09 Xy Inc SYSTEM FOR TREATING SPERMATIC CELL
ES2451765T3 (es) 2002-07-31 2014-03-28 Premium Genetics (Uk) Limited Sistema y procedimiento de clasificación de materiales usando orientación holográfica con láser
EP2275533B9 (en) 2002-08-01 2016-10-19 Xy, Llc Method of assessing sperm cells
US7855078B2 (en) 2002-08-15 2010-12-21 Xy, Llc High resolution flow cytometer
US7169548B2 (en) 2002-09-13 2007-01-30 Xy, Inc. Sperm cell processing and preservation systems
JP3983151B2 (ja) 2002-09-25 2007-09-26 ダイワ精工株式会社 魚釣用スピニングリール
US7201875B2 (en) 2002-09-27 2007-04-10 Becton Dickinson And Company Fixed mounted sorting cuvette with user replaceable nozzle
US20040061853A1 (en) 2002-09-27 2004-04-01 Blasenheim Barry J. Prism-based flow cytometry excitation optics
US6941005B2 (en) 2002-11-01 2005-09-06 Coulter International Corp. Monitoring and control of droplet sorting
CA2506935A1 (en) 2002-11-20 2004-06-03 University Of Virginia Patent Foundation Isolation of sperm cells from other biological materials using microfabricated devices and related methods thereof
WO2004059282A2 (en) 2002-12-19 2004-07-15 Monsanto Technology Llc Method and means for early detection of pregnancy in animals by combination testing
DK2959774T3 (da) 2003-03-28 2019-05-13 Inguran Llc Fremgangsmåde til kryokonservering af sædceller
BRPI0408852A (pt) * 2003-03-28 2006-04-04 Monsanto Technology Llc processo para a coloração de espermatozóide
WO2004104178A2 (en) 2003-05-15 2004-12-02 Xy, Inc. Efficient haploid cell sorting for flow cytometer systems
US20050011582A1 (en) 2003-06-06 2005-01-20 Haug Jeffrey S. Fluid delivery system for a flow cytometer
US7460223B2 (en) * 2003-09-19 2008-12-02 Applied Biosystems Inc. Inverted orientation for a microplate
BRPI0509485A (pt) 2004-03-29 2007-09-11 Monsanto Technology Llc suspensões de esperma para uso em inseminação
WO2005095960A1 (en) 2004-03-29 2005-10-13 Monsanto Technology Llc Use of a composition which regulates regulates oxidation/reduction reactions intracellularly and/or extracellularly in a staining or sorting process of spermatozoa
EP1769067A2 (en) 2004-07-22 2007-04-04 Monsanto Technology LLC Process for enriching a population of sperm cells
DK2884258T3 (da) 2004-07-27 2017-01-02 Beckman Coulter Inc Forbedring af flowcytometrisk diskrimination med computerimplementeret geometrisk transformation
US20060118167A1 (en) 2004-12-03 2006-06-08 Xy, Inc. Pressure regulated continuously variable volume container for fluid delivery
US20060147894A1 (en) 2004-12-30 2006-07-06 Vicam, L.P. Jacketed vessel for holding semen for sex biasing mammals through artificial insemination and systems and methods for enhancing the probability of sex biasing using the same
US7591064B2 (en) 2005-07-20 2009-09-22 Hitachi Global Storage Technologies Netherlands B.V. Method of fabrication for tape medium read head with unitary formation of multiple elements
US7618770B2 (en) 2005-07-29 2009-11-17 Xy, Inc. Methods and apparatus for reducing protein content in sperm cell extenders
CA2626518A1 (en) * 2005-10-19 2007-04-26 Scott Josephson Improved reproductive management

Also Published As

Publication number Publication date
HK1203831A1 (zh) 2015-11-06
EP2151243A3 (en) 2010-03-03
WO2005094852A3 (en) 2006-02-02
JP2009100752A (ja) 2009-05-14
EP2260854A1 (en) 2010-12-15
ES2397678T3 (es) 2013-03-08
JP2009082143A (ja) 2009-04-23
JP2007537727A (ja) 2007-12-27
NZ550198A (en) 2009-12-24
BRPI0509488A (pt) 2007-09-11
US7892725B2 (en) 2011-02-22
US20050214733A1 (en) 2005-09-29
CA2561519C (en) 2017-08-22
BRPI0509485A (pt) 2007-09-11
EP2801363A1 (en) 2014-11-12
AR048515A1 (es) 2006-05-03
CN102586179B (zh) 2015-04-08
AU2005229073B2 (en) 2010-08-19
WO2005095590A3 (en) 2005-12-08
NZ550196A (en) 2010-11-26
MXPA06011344A (es) 2006-12-15
EP1729789A2 (en) 2006-12-13
CA2561661C (en) 2015-11-24
CA2561661A1 (en) 2005-10-13
DK2801363T3 (en) 2018-05-28
EP2801363B1 (en) 2018-02-21
AU2005229073A1 (en) 2005-10-13
PT2151243E (pt) 2013-01-25
AR049259A1 (es) 2006-07-12
WO2005095590A2 (en) 2005-10-13
JP2007531523A (ja) 2007-11-08
AU2005228893A1 (en) 2005-10-13
HK1173745A1 (zh) 2013-05-24
CA2561519A1 (en) 2005-10-13
WO2005094852A2 (en) 2005-10-13
AU2005228893B2 (en) 2010-08-19
DK2151243T3 (da) 2013-01-28
CA2972254A1 (en) 2005-10-13
CN104974983A (zh) 2015-10-14
PL2151243T3 (pl) 2013-03-29
US7838210B2 (en) 2010-11-23
EP1730266A2 (en) 2006-12-13
EP2151243A2 (en) 2010-02-10
EP2151243B1 (en) 2012-10-24
US20050244805A1 (en) 2005-11-03
MXPA06011345A (es) 2006-12-15

Similar Documents

Publication Publication Date Title
CN102586179B (zh) 用于分选带有x或y染色体富集群的精子悬浮液
CN101238369B (zh) 调节细胞内和/或细胞外氧化/还原反应的组合物在精子染色或分选方法中的用途
EP2269617B1 (en) Process for enriching a population of sperm cells
CN1795004A (zh) 用于精子染色的方法
Emanuelli et al. Cumulus-oocyte interactions and programmed cell death in bovine embryos produced in vitro
CN101014700B (zh) 用于分选带有x或y染色体富集群的精子悬浮液
CN101048496B (zh) 富集精细胞群的方法
CN102643780B (zh) 富集精细胞群的方法
Aziz Effect of cumulus cells and β-mercaptoethanol supplement during in vitro maturation on bovine oocyte competency

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
REG Reference to a national code

Ref country code: HK

Ref legal event code: DE

Ref document number: 1173745

Country of ref document: HK

C14 Grant of patent or utility model
GR01 Patent grant
REG Reference to a national code

Ref country code: HK

Ref legal event code: GR

Ref document number: 1173745

Country of ref document: HK