CN101304683B - 以渐进增加的分辨率观察和分析一个或多个生物样品的方法和用于该方法的设备 - Google Patents

以渐进增加的分辨率观察和分析一个或多个生物样品的方法和用于该方法的设备 Download PDF

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CN101304683B
CN101304683B CN200680040695XA CN200680040695A CN101304683B CN 101304683 B CN101304683 B CN 101304683B CN 200680040695X A CN200680040695X A CN 200680040695XA CN 200680040695 A CN200680040695 A CN 200680040695A CN 101304683 B CN101304683 B CN 101304683B
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吉列尔莫·J·蒂尔尼
德维尔·叶林
本杰明·J·瓦科奇
梅利莎·苏特
布雷特·尤金·鲍马
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Abstract

提供了根据本发明的示范性实施例的方法、设备和装置，用于分析和/或图示解剖结构的至少一个部分。例如，将光转送到这样的部分，以便生成与该部分相关的第一信息。例如，将光提供到受检对象上或受检对象之内。接收第一信息，并且可以基于第一信息选择该部分的至少一个截面，以便生成第二信息。可以作为第二信息的函数而渐进修改该部分的显示的放大率。

Description

以渐进增加的分辨率观察和分析一个或多个生物样品的方法和用于该方法的设备
相关申请的交叉引用 
本申请基于并要求2005年9月29日提交的序列号为60/721,802的美国专利申请的权益，其整体内容通过引用结合于此。 
技术领域
本发明涉及用于以渐进增加的分辨率观察和分析一个或多个生物样品和解剖结构的方法和装置。 
背景技术
放射技术如X射线计算机断层扫描(“CT”)、磁共振成像(“MRI”)和超声波能够使得器官水平的人体病变的非侵入可视化成为可能。尽管这些医疗器械可能能够识别大尺度的病变，但是癌症的诊断能够需要微观结构的评估，这超出了传统成像技术的分辨率。因此，诊断可能需要活组织检查和病理组织学检查。因为癌症前期的生长和早期癌症常常以微观尺度出现，所以它们能够为识别和诊断提供显著的挑战。这些病变的传统筛选和监视依赖于苏木精和伊红(“H&E”)染色玻片的无制导的活组织检查和形态分析。尽管这种方法可以被认为是用于微观诊断的当前标准，但是它需要从患者去除组织并且需要显著的处理时间以生成玻片。更重要地，组织病理学固有地为点采样技术；常常只有非常小的一部分的患病组织能够被切除，并且病理学家常常可能检查小于1％的活组织检查样品。 
可以优选的是，从活人患者的整个器官或生物系统中获得微观诊断。然而，合适成像技术的缺乏极大地限制了用于筛选肿瘤前的状况(例如转化)和发育异常的选项。另外，无能力现场识别发育异常和恶性肿瘤的区域已导致诸如像前列腺、结肠、食道和膀胱等等的随机活组织检查的筛选过程，这是高度不希望的和杂乱无章的。当前涉及冷冻切片实验室的许多诊断任务如外科肿瘤边界的描绘可以通过诊断医疗器械来改进，该诊断医 疗器械能够以微观尺度迅速成像大的组织体积。能够填补病理学和放射学之间的这个空白的技术将会对患者管理和卫生保健具有重大利益。 
已进行了技术进步以增加非侵入成像技术诸如像微CT、微PET和磁共振成像(“MRI”)显微的分辨率。通过这些技术已实现了接近20μm的分辨率，但是基本的物理限制仍然阻止它们应用于患者。对于非切除的组织病理学诊断，现场进行的微观光学活组织检查技术近来已取得进展。反射共焦显微(“RCM”)可以特别好地适合于对患者的非侵入显微镜检查，因为它能够测量微观结构而没有组织接触，并且不需要管理外在造影剂。RCM能够抑制离焦光，并且检测选择性地起源于组织内单平面的背散射的光子。例如通过在平行于组织表面的平面内迅速扫描电磁辐射的聚焦束，能够实施RCM，获得组织的横断或表面图像。可以用在RCM中的大数值孔径(NA)能够获得非常高的空间分辨率(1-2μm)，使得亚细胞结构的可视化成为可能。然而，高NA成像对随着光传播通过不均匀的组织而出现的像差特别敏感。而且，使用RCM的高分辨率成像典型地被限制到大约100-400μm的深度。 
RCM已被广泛地展示为用于表皮组织的可行成像技术。内窥镜共焦显微系统的开发已变得更加困难，这至少部分地归因于涉及使扫描显微镜小型化的重要技术挑战。将共焦显微镜检查的概念直接应用于内窥镜检查的一个主要障碍是机构的操纵，该机构用于在小直径软质探针的远端快速扫描聚焦束。已提议了多种方法来应对这个问题，包括远端微机电系统(“MEMS”)束扫描装置的使用和单模光纤束的近端扫描。而且，RCM可以提供仅在离散位置处的微观图像——“点采样”技术。如当前实施的那样，点采样是RCM所固有的，因为它具有有限的视场，其可以与切除活组织检查的视场相当或比它小，并且对于全面的大视场显微镜检查，成像速率太慢。 
使共焦显微镜检查适用于内窥镜应用的另一个挑战包括可以用于光学切片的高NA物镜的小型化。通过提供例如梯度指数透镜系统、双轴物镜或定制设计的小型化物镜，可以实现这样的小型化。例如，使用耦合到小型化物镜的光纤束，可以在体内获得子宫颈上皮的形态的详细图像，并且使用商用仪器如可以从例如Olympus Corp.(奥林巴斯公司)和Pentax/Optiscan获得的商用仪器，可以得到结直肠病变的基于荧光的图像。 
尽管有这些进展，仍然需要提供方法和装置，该方法和装置能够解析(例如从大的表面面积乃至可能整个器官获得的以细胞水平、结构水平或 两者提供的)数据，以便它可以以及时、准确的方式被适当地解释。事实上，这种数据的量很大并且难以同时观察这样的数据，因而这样的方法和装置将会对于其观察和分析有益。 
发明内容
本发明的目的之一是要克服现有技术系统的某些缺陷和缺点(包括上面在此描述的那些)，并且提供用于以渐进增加的分辨率观察和分析一个或多个生物样品和解剖结构的方法和装置的示范性实施例。这样的示范性方法和装置能够和数据的目视检查一起使用，或者通过数据的自动处理过程使用，以引导最有可能包含异常和/或不健康组织的区域的可视化。 
因此，提供了根据本发明的示范性实施例的方法、设备和装置，它们可以分析和/或图示解剖结构的至少一个部分。例如，在这样的示范性实施例中，将光转送到这样的部分，以便生成与该部分相关的第一信息。例如，将光提供到受检对象上或受检对象之内。接收第一信息，并且可以基于第一信息选择该部分的至少一个截面，以便生成第二信息。可以作为第二信息的函数而渐进修改该部分的显示的放大率。 
在本发明的进一步的示范性实施例中，(例如在解剖结构之内)能够修改所述至少一个部分的位置和/或深度的显示。第二信息能够与在这样的部分之内提供的区域相关联，和/或可以通过用户选择该区域来获得。该选择能够通过处理装置自动进行而不用从用户输入。可以确定所述至少一个部分之内的异常性区域，并且处理装置可以进行选择和修改，以便显示异常性的至少一个截面。使用处理装置能够确定该部分之内的异常性区域以便生成第三信息，并且该选择能够作为第三信息的函数而被用户和/或处理装置执行。 
根据本发明的再一个示范性实施例，第一信息能够与该部分的二维、三维、四维或更多维表示相关联。进一步，该部分可以具有大于1mm2 和/或10mm2的面积。该部分的显示的截面可以具有小于1cm2、1mm2和/或100μm2的面积。第一信息能够与共焦显微镜检查过程、谱编码共焦显微镜检查过程、光学相干断层扫描过程和/或光学频域干涉量度过程相关联。装置位于解剖结构之内，以便将光提供给该部分。 
结合所附的权利要求书，当阅读以下对本发明实施例的详细描述时，本发明的其它特征和优点将会变得明显。 
附图说明
结合示出了本发明的示意性实施例的附图，从以下详细描述中，本发明的进一步的目的、特征和优点将会变得明显，其中： 
图1是示范性谱编码共焦显微镜检查(SECM)系统的示意性图示； 
图2A是使用单模源和单模检测(SM-MM)配置离组织表面100μm在体外获得的猪肠上皮的示范性SECM图像； 
图2B是使用单模源和多模检测(SM-MM)配置获得的猪肠上皮的另一个示范性SECM图像； 
图2C是猪肠上皮的SECM图像的放大图； 
图3A是在以50μm的成像深度压缩肠壁之后在体外获得的猪肠上皮的示范性SECM图像； 
图3B是在以100μm的成像深度压缩肠壁之后在体外获得的猪肠上皮的示范性SECM图像； 
图4是示范性SECM设备的示意性图示； 
图5是USAF图表的示范性SECM图像； 
图6A是以1x的放大率显示的基于取自镜头纸样品的数据的示范性SECM图像； 
图6B是以4.5x的放大率显示的基于取自镜头纸样品的数据的示范性SECM图像； 
图6C是以16.7x的放大率显示的基于取自镜头纸样品的数据的示范性SECM图像； 
图6D是以50x的放大率显示的基于取自镜头纸样品的数据的示范性SECM图像； 
图6E是以125x的放大率显示的基于取自镜头纸样品的数据的示范性SECM图像； 
图7是在5个不同焦点位置从镜头纸样品获得的一系列示范性SECM数据以及通过组合5个各自图像中的数据生成的组合图像； 
图8A是以1x的放大率显示的基于取自猪肠组织片段的数据的示范 性SECM图像； 
图8B是以4x的放大率显示的基于取自猪肠组织片段的数据的示范性SECM图像； 
图8C是以20x的放大率显示的基于取自猪肠组织片段的数据的示范性SECM图像； 
图8D是以40x的放大率显示的基于取自猪肠组织片段的数据的示范性SECM图像； 
图9A是在体内的猪食道的微观图像的前视图和侧视图，其使用根据本发明的方法和装置的示范性实施例示出了黏膜下层之内的脉管网络，而没有使用图像增强剂或外在的造影剂； 
图9B是通过图9A中图示的位置处的食道壁的纵向横截面的微观图像的侧视图； 
图9C是A中图示的位置处的打开横断截面的侧视图； 
图9D是图9C中图示的图像的选择截面的展开图的侧视图； 
图9E是从对应于图9D中图示的图像的解剖区域获得的代表性组织截面的示范性图像； 
图10是本发明的用于渐进变焦到解剖结构的微观数据集中的方法的示范性实施例的流程图； 
图11是使用光学相干断层扫描(“OCT”)技术获得的食道黏膜的一系列示范性图像，展示了用于识别与Barrett的食道和腺癌相比较的正常鳞状黏膜的自动处理过程的示范性实施例的实施； 
图12是使用OCT技术获得的动脉粥样硬化斑的一组示范性图像，其已被处理以识别巨噬细胞密度；以及 
图13是根据本发明的方法的另一个示范性实施例的流程图，该方法用于基于经由信号处理技术获得的结果，渐进变焦到解剖结构的微观数据集，以自动识别可以以高放大率观察的所关心区域。 
贯穿附图，除非另外声明，否则相同的标号和字符用于指示图示实施例的相同特征、元件、部件或部分。此外，虽然现在将参考附图详细地描述本发明，但是这将结合示意性实施例进行。能够对描述的实施例进行改变和修改，而不认为脱离了如所附权利要求所限定的本发明的真实范围和 精神。 
具体实施方式
根据本发明的示范性实施例，能够提供根据本发明的示范性实施例的方法和装置，用于以渐进增加的分辨率观察和分析一个或多个生物样品和解剖结构。这样的示范性方法和装置能够和数据的目视检查一起使用，或者通过数据的自动处理过程使用，以引导最有可能包含异常和/或不健康组织的区域的可视化。 
在图1中示出了示范性SECM技术。来自可以位于探针远端的单模光纤100的输出能够被准直透镜110校准，然后照射色散光学元件(诸如像透射衍射光栅120)。物镜130然后能够将每个衍射波长聚焦到样品之内的不同空间位置，导致横断线状焦点140，其中线上的每个点用不同的波长表征。在从可以例如是生物组织的样品反射之后，光信号可以被衍射元件120重新组合并由单模光纤100收集。单模光纤100的核心孔径可以提供空间滤波机制，其能够滤去散焦光。在探针外部(并且任选地在系统控制台之内)，返回的光的光谱可以被测量并被转换成作为样品之内横向位移的函数的共焦反射。能够快速执行谱解码。这样一来，通过相对缓慢和直接的机械动作，就能够完成通过在正交于线状焦点的方向上扫描射束而产生的图像。 
SECM技术可以允许使用内窥镜RCM，并且能够通过使用高速线性CCD摄像机以极高速率提供图像数据。商业上可用的线性CCD阵列能够以大于大约每秒60兆像素的速率获得数据。当结合到SECM分光计中时，这些阵列能够以大约10倍于典型视频速率并且一直到100倍于某些内窥镜RCM技术的速度产生共焦图像。典型SECM系统的快速成像速率和光纤设计允许通过内窥镜探针的全面的大面积显微镜检查。 
使用光学相干断层扫描(“OCT”)的技术及其变体可以用于全面的构造筛选。获取波长域中的而不是时域中的OCT信号，能够提供成像速度的数量级改善，同时维持良好的图像质量。使用谱域OCT(“SD-OCT”)技术，通过检测组织样品和基准之间的谱分辨干涉，能够在生物组织中实施高分辨率测距。因为SD-OCT系统能够使用与SECM系统相同的高速线性CCD，所以它们也能够以每秒60兆像素捕捉图像，这近似于两个数量级的传统时域OCT(“TD-OCT”)系统的速率。使用这种获取速率和 分辨率，SD-OCT系统能够在临床环境中以构造水平提供全面的体积显微镜检查。 
SD-OCT和SECM系统提供的信息可以是互补的，并且使用这两种技术的混合平台能够提供可能对于准确诊断所必需的有关组织的构造和细胞结构的信息。尽管不同技术的结合典型地需要大范围的工程并且可能使性能折衷，但是SECM和SD-OCT系统能够共享关键的部件，并且能够提供高性能多模态的系统，而不会显著增加单个系统的复杂性或成本。 
根据本发明的某些示范性实施例的SECM系统能够使用波长扫描1300nm的源和单元件光检测器来获得作为时间函数的谱编码信息。使用这个系统，能够以在400μm视场(“FOV”)之上具有高横向(1.4μm)和轴向(6μm)分辨率的高达约30帧/秒的速率获取图像。用高速系统在体外成像新近切除的猪十二指肠片段的图像，以展示SECM系统识别亚细胞结构的能力，该亚细胞结构可以在专化肠上皮化生(“SIM”)即BE的化生变化中发现。 
图2A-2C描绘了在体外获得的猪肠上皮的示范性SECM图像，其使用了两种成像模式和相应的光纤配置：具有单模检测的单模照射(“SM-SM”)以及具有多模检测的单模照射(“SM-MM”)。图2A中的SM-SM图像示出了使用单模源和单模检测的距离组织表面100μm的上皮结构。使用具有1∶4的核心-孔径比的单模源和多模检测(SM-MM)获得的示出在图2B中的相同组织区域的图像，因为斑点噪声减少而显得更加平滑并且可以更加容易地解释。图2C是图2B中示出的图像的放大图，其示出了包含不良反射的核心(例如固有层或“lp”)和较高散射的柱状上皮的绒毛的证据。在柱状细胞的底部可见的明亮图像密度，其与核一致(用箭头指示)，在图2C中是明显的。 
使用膨胀气囊，能够将使用OCT技术在体内成像的食道壁的厚度减少例如大约二分之一。图2A-2C中示出的猪肠样品被减少相同的量，并且使用SECM技术观察到的亚细胞特征被很好地保存。图3A和3B分别示出了在50μm和100μm的深度获得的该变薄样品的图像。 
商用800nm激光扫描共焦显微镜的穿透深度被观察到与用1300nmSECM系统获得的穿透深度相比减少了大约20％。减少的穿透可能是较短波长源的散射增加的结果。这样一来，使用840nm源的SECM系统就可以提供足够的穿透，以识别例如肠上皮的亚细胞结构。 
在图4中示意性地图示了被配置成提供全面SECM图像的根据本发明的某些示范性实施例的设备。这个示范性设备被配置成从圆柱形样品获得图像，该圆柱形样品具有2.5cm的长度和2.0cm的直径，这近似为远端食道的尺度。具有以800nm为中心的波长和45nm的带宽的光纤耦合的2.0mW超辐射二极管200(QSSL-790-2，qPhotonics，Chesapeake，VA)，被配置成照射50/50单模光纤分束器405。被传输通过分束器的一个端口的光被准直仪410校准，并且被传输通过光纤412到达聚焦设备415并到达光栅透镜对，该光栅透镜对包括：光栅420(1780lpmm，Holographix，LLC，Hudson，MA)；以及350230-B非球面透镜425(Thor Labs，Inc.，Newton，NJ)，其具有4.5mm的焦距f、5.0mm的通光孔径和0.55的NA。这种布置能够在圆柱形样品的内表面上产生聚焦的谱编码的斑点430的500μm纵向线性阵列或线。光栅透镜对通过壳体440附着到马达435(1516SR，15mm直径，MicroMo Electronics，Inc.，Clearwater，FL)的轴。随着马达435旋转，跨越圆柱形样品的内圆周扫描谱编码线。使用计算机控制的线性级445(Nanomotion II，2.5cm range，Melles Griot，Rochester，NY)，在马达435的旋转期间沿着圆柱形样品的纵轴平移马达435、壳体440和光栅透镜对。这个过程产生了圆柱形样品的整个内表面的螺旋扫描。 
从样品反射的光被传输返回通过光学系统进入单模光纤412，并且被光纤412提供给分光计450和线性CCD 455，该线性CCD 455包括2048个像素并且具有30kHz的线速率(Basler L104K，Basler VisionTechnologies，Exton，PA)。计算机460用于存储、分析和显示分光计450和CCD 455提供的图像数据。每周马达旋转大约有60,000点(以0.5Hz或30rpm)被数字化，以实现1.0μm的圆周采样密度。马达的纵向速度为0.25mm/s，并且圆柱形样品的一个完整扫描所需的时间为100秒。 
光栅透镜对上的准直束的1/e2直径为4.0mm。结果，这个示范性设备的有效NA近似为0.4，这对应于近似1.2μm的理论斑点直径和近似2.5μm的共焦参数。在没有光学像差的系统中，样品上的理论谱分辨率可以为 
Figure S200680040695XD00081
其能够跨越谱编码线430得到高达近似630个可分辨点。检测臂中的分光计450被设计成超过探针的预测谱分辨率。 
在图5中示出了使用这种设备获得的1951 USAF分辨率图表的SECM扫描。被分开2.2μm的这个图中的最小条被分辨。使用扫描通过焦点的镜获得的横断线扩展函数半高宽(“FWHM”)和轴向FWHM函 数分别被测量为2.1μm和5.5μm。观察到视场大约为500μm。这些测量结果略微低于相应的理论值，这可能归因于光路中的像差。这些实际的参数表明，在此描述的示范性设备能够提供足够的分辨率以用于生物组织中的共焦显微镜检查。 
在图6中示出了用于2.5cm体模样品的完全回拉图像的SECM图像数据。在生成这些显示的图像之前，将极坐标转换成直角坐标。使用附着到2.1cm内径的特氟隆(Teflon)管的内表面的镜头纸来制作体模样品。在图6A示出的低放大率图像中，能够观察到纸的包括褶皱和空隙的宏观结构。可见的圆周条纹可能起因于存在于谱编码线的末端处或其附近的较低的谱功率和透镜像差。如图6B-6E所示，在以较高的放大率提供的这个数据集的区域中，能够清楚地分辨各个纤维和纤维微观结构。 
通过调整图4A中的聚焦设备415，在120μm的范围的5个离散聚焦深度处获取仿真样品的圆柱形二维(“2D”)图像。图7中示出的这5个图像710-750然后被求和以产生综合图像760，其展示了体模样品表面的几乎完整的覆盖。 
由于缺少用于光学扫描头的定心设备，使用如在此描述的SECM设备来成像生物样品能够被复杂化。为了提供进一步的改进以便生成宽视场显微图像和数据，在2.0cm直径的透明圆柱之上放置猪肠的样品。在图8A中示出了1秒内获取的这个样品的360°扫描。成像的组织仅在圆柱形扫描的一个区段中出现，因为探针未被置于中心，并且样品没有完全缠绕在圆柱周围。图8B-8D示出了这个组织样品的一系列放大区域。图8B中示出的图像是图8A中的打点矩形框出的1.5cm区段的扩展。类似地，图8C中的图像表示了图8B中框出的矩形的扩展，而图8D中的图像则表示了图8C中框出的矩形的扩展。图8B中的组织的放大图像暗示了腺结构。图8C-8D中的放大图像展示了绒毛和核特征，它们类似于如图2和3所示的使用1300nm SECM系统观察到的绒毛和核特征。图8A中的SECM扫描的其它区域示出了伪像(artifact)，包括来自透明圆柱的镜反射和全信号丢失，这两者都可能起因于聚焦SECM束的不适当定位。 
对患者实施全面共焦显微镜检查提出了多种技术挑战。这样的挑战可能包括例如增加成像速率、使探针光学部件和机械部件小型化、结合定心机构以及实施用于动态改变焦平面的技术。 
与在上文描述的示范性系统相比，能够将SECM系统的图像获取速度改进为该示范性系统的例如大约2-4倍。通过提供某些修改能够实现这 样的改进。例如，更高功率半导体光源(诸如像超辐射二极管T-840HP：25mW，840nm，100nm谱带宽)能够提供1000个谱可分辨点。光功率的增加能够改进灵敏度，并且较大的带宽可以拓宽视场，使得可以以近似两倍的速度扫描SECM束。而且，使用光环行器诸如像OC-3-850(Optics for Research，Caldwell，NJ)能够增加被传递到探针和从探针收集的光的效率。使用更快更灵敏的线性CCD诸如像具有2048个像素和60kHz读出速率的AVIIVA M4-2048(Atmel Corporation)能够提供两倍的数据获取速度和用于生成图像数据的波长范围的改进的谱响应。还可以通过使用例如Camera Link(摄像机链接)接口来改进性能，所述Camera Link接口能够以近似120MB/s的速率从摄像机向用于存储的硬盘驱动器阵列传送数据。 
被理解为指的是最小可检测反射率的灵敏度是影响共焦图像质量和穿透深度的系统参数。当使用近红外RCM技术时，能够从一直到近似300μm的深度处的皮肤反射入射光的一小部分，其可能近似为10-4到10-7 。基于在此描述的根据本发明的某些示范性实施例的示范性系统中使用的物镜的NA以及皮肤比非角质化上皮黏膜更加显著地可以使光衰减的观察，在此描述的示范性SECM探针物镜可以收集近似3×10-4到3×10-7 的从组织之内深处反射的照射光。25mW光源可以被分成例如近似1000个独立的射束。最大双程插入损耗被估计为近似10dB(6dB来自探针，4dB来自光纤和分光计)。基于这些估计的参数，对于每线集成周期，从而可以用近似50到50,000个光子/像素来照射阵列中的每个像素。 
使用多模检测技术，可以实现10倍的信号增益，使得对于这样的配置每次扫描有近似500到500,000光子/像素。如果信号在以近似240个光子发生的暗电流波动之上，则例如Atmel AVIIVA M4摄像机上的单个像素能够可靠地检测光。如果这个装置在这些波长处具有近似50％量子效率，则能够以每次扫描有近似480光子/像素产生最小可检测信号。基于这些近似，Atmel摄像机可以具有足够的灵敏度以允许较深的组织深度处的SECM成像。通过使用多模光纤来收集或者通过增加源功率，能够实现对预测的最小反射率进行的量子噪声限制检测。 
根据本发明的一个示范性实施例，能够提供用于导航、分析和显示来自解剖结构的大的微观数据集的方法和装置。 
图9A-9E图示了使用全面的显微镜检查与本发明的方法和装置的示范性实施例获得的体内猪食道的各种图像。这些示范性图像由图4中示出 的计算机460(例如个人计算机、微型计算机等等)生成，或者由别的处理装置生成，所述计算机460或处理装置可以(例如通过软件)被配置成将这样的图像转发到图4的显示器470或别的输出装置。另外，计算机460控制图4的示范性系统的各种部件(例如马达435、线平移器445、聚焦设备415等等)，以自动地和/或在人工控制之下聚焦在解剖结构的各个区域上，这将使得能够导航、分析和显示与解剖结构相关联的大的微观数据集。 
例如，图9A分别示出了在体内的猪食道的微观图像的前视图和侧视图900、905，其使用根据本发明的方法和装置的示范性实施例提供了黏膜下层之内的脉管网络，而没有使用图像增强剂或外在的造影剂。事实上，可以对图9A的例如14GB容量的数据集进行再现和降采样，以用于在任意的取向和透视图中呈现。在没有使用这样的图像增强剂或外在的造影剂的情况下，示出了黏膜下层之内的脉管网络。使用为(一个或多个)这样的示范性任务配置的计算机460和图4的系统的其它部件，能够在体积图像上定位横截面图像，用于更高分辨率观察。 
图9B示出了通过图9A中图示的位置处的食道壁的纵向横截面的微观图像的侧视图910。例如，这个图像910以上皮在顶部的方式反转；尺度：水平45mm，垂直2.6mm。在原始数据中，能够观察到对应于心跳运动的周期性垂直偏移。表面对准过程的示范性实施例用于减少这种伪像而只剩余垂直条带，该剩余的垂直条带仍然可以以对应于90次/分钟的心率的300微米的周期被观察到。相邻A线之间的示范性纵向间距被示出为32μm。 
图9C示出了图9A中图示的位置处的打开横断截面(例如圆柱坐标r和θ被映射到垂直和水平坐标)的侧视图920。例如，图示的示范性尺度如下：57mm-水平，2.6mm-垂直。图9B和9C都图示了通过整个食道壁的成像，并且能够识别鳞状上皮(e)、固有层(lp)、黏膜肌层(mm)、黏膜下层(s)和固有肌层(mp)。图9D示出了有助于这样的识别的图9C中图示的图像的选择截面的展开图的侧视图930。图9E示出了从对应于图9D中图示的图像的解剖区域获得的代表性组织截面(H&E染色)的示范性图像。 
例如，图10描绘了流程图，其描述了能够使用图4中示出的计算机460来执行的用于以渐进更高的分辨率分析和/或观察数据集的根据本发明的方法或过程的示范性实施例。具体地，在步骤1000中，在组织的大 面积之上，或者从组织或其中的器官的体积中，可以获取具有小于10μm的分辨率的微观数据集。然后能够以如下的表达(在步骤1010中)将数据格式化：可以图示整个数据集或数据集的一部分的低放大率或低倍视图。在步骤1020中，用户可以观察数据集，并且使用计算机接口选择(a)矩形区域、(b)点、(c)任意成形区域和/或(d)深度，以目视更高放大率视图。在步骤1020中观察新的区域，并且用户在数据集之内(a)选择另一个区域、(b)在点处放大、(c)缩小、(d)在三维中平移当前视图和/或(e)改变观察的深度位置。 
重复图10中图示的整个示范性过程，直到所关心的一个或多个区域被可视化地识别为止。用户以任何放大率或视图来选择不同的图像，以存储用于稍后的检查。在示范性导航过程期间还能够对每个单独视图进行标记。不同放大率/位置的各种区域/图像能够被标上书签，以便用户能够在随后的导航会话期间返回到相同的区域/图像。图6A-6E、7和9A-9E图示了观察大面积微观数据集时的渐进放大率的例子。 
在此描述的导航过程的示范性实施例通过计算机460来实施，并且还利用各种处理技术辅助用户确定各种区域以放大和观察样品及其不同的部分和区域。例如，图11描绘了使用光学相干断层扫描(“OCT”)技术获得的食道黏膜的一系列示范性图像，展示了用于经由OCT图像空间频率的分析而识别与Barrett的食道和腺癌相比较的正常或良性鳞状黏膜的自动处理过程的示范性实施例的实施。 
如图11所示，示出了不同病状的OCT图像1100、1110和1120以及空间频率分布1105、1115和1125。鳞状上皮1100(SE)具有垂直空间频率(见面板1105中的箭头1007)，其对应于在SIM中可能不存在的水平层。与SIMND 1110和1115相对照，在腺癌(CA)1120的示范性OCT图像及其相应的空间频率1125中示出了广泛变化的空间频率分布。图12描绘了通过确定归一化的标准偏差参数(NSD)而从动脉粥样硬化斑1200的OCT图像中获得的巨噬细胞含量1210的图示。使用色表1220，可以获得巨噬细胞的密度并将其显示为图像。 
这些以及其它的示范性图像处理分析过程和步骤能够被应用于微观数据集并用于突出潜在疾病的区域，用于随后的定向导航。图13描绘了用于导航和评估微观图像数据集的根据本发明的方法和过程的示范性实施例的流程图。在这个示范性方法/过程中，在步骤1300中获得微观数据集，其优选地具有在组织的大面积之上或者从组织或其中的器官的体积中 获取的小于10μm的分辨率。然后在步骤1310中通过处理装置(例如使用计算机460)自动地处理数据，以识别包含疑似疾病区域或相反地疑似不包含疾病区域(亦即健康部位)的区域/位置。在步骤1320中，不健康区域通过使用颜色或其它标记方法而被表达，然后在步骤1330中以整个微观数据体积的低放大率而被观察。 
用户然后能够由处理数据及其表达引导，在步骤1340中选择区域来观察。然后用户可以观察数据集，并且使用计算机接口来选择(a)矩形区域、(a)点、(c)任意成形区域和/或(d)深度，以目视更高放大率视图。可以观察新的区域，并且用户(或计算机460)可以在数据集之内手动或自动地(a)选择另一个区域、(b)在点处放大/缩小(步骤1350)、(d)在三维中平移当前视图和/或(e)改变观察的深度位置。进一步，在步骤1360中，用户观察新近图示的区域。可以重复示范性的方法/过程，直到所关心的(一个或多个)区域被可视化地识别为止。用户或计算机460可以选择任何放大率或视图的不同图像，以存储用于稍后检查。在示范性导航过程期间还可以对每个单独视图进行标记。不同放大率/位置的各种区域/图像可以被标上书签并存储，以便用户能够在随后的导航会话期间返回到相同的区域/图像。 
前述仅仅表明了本发明的原理。考虑到在此的教导，对所描述的实施例的各种修改和变更对本领域技术人员而言将会是明显的。事实上，根据本发明的示范性实施例的装置、系统和方法，可以与任何OCT系统、OFDI系统、SD-OCT系统或其它成像系统一起使用，并且例如与2004年9月8日提交的国际专利申请PCT/US2004/029148、2005年11月2日提交的美国专利申请No.11/266,779和2004年7月9日提交的美国专利申请No.10/501,276中描述的那些一起使用，这些专利申请的整体内容通过引用结合于此。这样一来，将会意识到的是，本领域技术人员将会设计众多的系统、装置和方法，它们尽管没有在此明确地示出或描述，但却体现了本发明的原理，并从而处在本发明的精神和范围之内。另外，就上面尚未明确地将现有技术知识通过引用结合于此而言，明确地以其整体结合于此。上面引用在此的所有公布都以其整体通过引用结合于此。 

Claims (22)

1.一种用于分析和/或图示解剖结构的至少一个部分的方法，包括：
将光转送到所述至少一个部分以便生成与所述至少一个部分相关的第一信息，其中，位于包含所述解剖结构的受检对象之内的生成装置将所述光提供到所述受检对象上或所述受检对象之内，所述生成装置生成所述第一信息，所述第一信息与图像相关联；
接收所述第一信息，并且基于所述第一信息选择所述至少一个部分的至少一个截面，以便生成第二信息；以及
渐进地修改作为所述第二信息的函数的所述至少一个部分的显示的放大率。
2.根据权利要求1所述的方法，其中，所述修改步骤包括修改所述至少一个部分的位置的显示。
3.根据权利要求1所述的方法，其中，所述修改步骤包括修改所述至少一个部分在所述解剖结构之内的深度的显示。
4.根据权利要求1所述的方法，其中，所述第二信息与所述至少一个部分之内提供的区域相关联。
5.根据权利要求1所述的方法，其中，通过用户选择所述区域来获得所述第二信息。
6.根据权利要求1所述的方法，其中，所述选择步骤通过处理装置自动执行而不用从用户输入。
7.根据权利要求1所述的方法，进一步包括确定所述至少一个部分之内的异常性区域，并且其中处理装置执行所述选择步骤和修改步骤，以便显示所述异常性的至少一个截面。
8.根据权利要求1所述的方法，进一步包括使用处理装置确定所述至少一个部分之内的异常性区域，以便生成第三信息，并且其中所述选择步骤作为所述第三信息的画数而被用户执行。
9.根据权利要求1所述的方法，进一步包括使用处理装置确定所述至少一个部分之内的异常性区域，以便生成第三信息，并且其中所述选择步骤作为所述第三信息的函数而被所述处理装置执行。
10.根据权利要求1所述的方法，其中，所述第一信息与所述至少一个部分的二维表示相关联。
11.根据权利要求1所述的方法，其中，所述第一信息与所述至少一个部分的三维表示相关联。
12.根据权利要求1所述的方法，其中，所述第一信息与具有多于三维的所述至少一个部分的表示相关联。
13.根据权利要求1所述的方法，其中，所述至少一个部分具有大于1mm2的面积。
14.根据权利要求1所述的方法，其中，所述至少一个部分具有大于10mm2的面积。
15.根据权利要求1所述的方法，其中，所述至少一个部分的显示的截面具有小于1cm2的面积。
16.根据权利要求1所述的方法，其中，所述至少一个部分的显示的截面具有小于1mm2的面积。
17.根据权利要求1所述的方法，其中，所述至少一个部分的显示的截面具有小于100μm2的面积。
18.根据权利要求1所述的方法，其中，所述第一信息与以下中的至少一个相关联：
共焦显微镜检查过程；
谱编码共焦显微镜检查过程；
光学相干断层扫描过程；以及
光学频域干涉量度过程。
19.根据权利要求1所述的方法，进一步包括在所述解剖结构之内提供装置，以便将所述光提供给所述至少一个部分。
20.根据权利要求1所述的方法，其中，所述至少一个部分是血管。
21.一种用于分析和/或图示解剖结构的至少一个部分的设备，包括：
至少一个第一装置，其位于包含所述解剖结构的受检对象之内并且配置成将光转送到所述至少一个部分上或所述受检对象之内，以便生成与所述至少一个部分相关的第一信息，所述第一信息与图像相关联；
至少一个第二装置，其配置成接收所述第一信息，并且基于所述第一信息选择所述至少一个部分的至少一个截面，以便生成第二信息；以及
至少一个第三装置，其配置成渐进地修改作为所述第二信息的函数的所述至少一个部分的显示的放大率。
22.根据权利要求21所述的设备，其中，所述至少一个部分是血管。
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