WO1996018099A1 - Arrangement in a gel electrophoresis apparatus - Google Patents
Arrangement in a gel electrophoresis apparatus Download PDFInfo
- Publication number
- WO1996018099A1 WO1996018099A1 PCT/SE1995/001482 SE9501482W WO9618099A1 WO 1996018099 A1 WO1996018099 A1 WO 1996018099A1 SE 9501482 W SE9501482 W SE 9501482W WO 9618099 A1 WO9618099 A1 WO 9618099A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- plates
- fluorophore
- optical filter
- gel
- laser
- Prior art date
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44717—Arrangements for investigating the separated zones, e.g. localising zones
- G01N27/44721—Arrangements for investigating the separated zones, e.g. localising zones by optical means
Definitions
- the present invention relates to an arrangement in a gel electrophoresis apparatus having a laser which is adapted to emit a laser beam of a predetermined wavelength to excite at least one substance which is labelled with a fluorophore, and which is caused to migrate along a gel which is provided between two transparent plates, a detector which is located close to one of the plates to detect the fluorescence emitted upon excitation of the fluorophore, and an optical filter which is located between said one of the plates and the detector, and which is adapted to block light of said predetermined wavelength.
- fluorophore i.a. a carbocyanine based dye marketed under the trademark Cy5 by Biological Detection Systems, Inc., Pittsburg, PA, U.S.A.
- the dye Cy5 can be excited by means of a laser, e.g. a helium-neon laser, operating at a wavelength of 633 n , or a laser diode of a wavelength between 630 and 650 n .
- the fluorescence emitted upon excitation of the dye Cy5 will have a wavelength of between 650 and 720 nm with a maximum at about 670 nm.
- the object of the invention is to bring about an arrangement which makes it possible to eliminate the effect of this disturbing source.
- This is attained according to the invention in that the optical filter is adapted to block also the intrinsic fluorescence of the plates.
- Fig. 1 is a schematic cross-sectional view of a portion of an embodiment of a gel electrophoresis apparatus with an arrangement according to the invention
- Fig. 2 is a schematic diagram of fluorescence and filter characteristics in the embodiment shown in Fig. 1.
- Fig. 1 denotes an electrophoresis gel provided between two plates 2 and 3.
- the plates 2 and 3 are transparent and are preferably made of glass but can also be made of plastic.
- At least one substance 4, labelled with a fluorophore, e.g. the carbocyanine based dye Cy5, mentioned in the introduction, is adapted to migrate in a manner not shown under the influence of an electric field.
- a laser not shown, e.g. a helium-neon laser or a laser diode, is provided to supply into the gel 1, through one of the lateral edges of the gel 1, a laser beam of a predetermined wavelength to excite the fluorophore of the substance 4 migrating along the gel 1.
- Fig. 1 the substance 4 is schematically shown in the position where its fluorophore has been excited by the laser beam, not illustrated, going across the gel 1.
- a detector 5 e.g. a photo diode
- An optical filter 6 is provided between the detector 5 and the plate 3. According to the invention, the optical filter 6 is adapted to block not only light of the wavelength of the laser beam but also light originating from the intrinsic fluorescence of the glass plates 2 and 3 upon the excitation by the laser light.
- reference numeral 7 designates laser wavelength
- 8 is the characteristic of an optical filter according to the invention
- 9 designates fluorescence from a fluorophore
- 10 is fluorescence from glass plates.
- the fluorescence from the fluorophore detected by the detector 5 will exhibit a maximum at a wavelength of about 670 nm, while the maximum of the intrinsic fluorescence of the glass plates 2 and 3 will appear at a wavelength of about 770 nm.
- the optical filter 6 is designed as a band pass filter to block light of the laser wavelength as well as light originating from the intrinsic fluorescence of the glass plates 2 and 3.
- a number of detectors 5 and a number of optical filters 6 are provided to detect fluorophore labelled substances migrating along a corresponding number of parallel paths in the gel 1.
- the optical filter 6 is preferably dimensioned in such a manner that it exhibits a pass band characteristic having a center wavelength of 680 ⁇ 10 nm, a half-width of 40 ⁇ 5 nm and an attenuation which is greater than OD5 between 400 and 640 nm, and between 720 and 800 nm, and greater than OD3 between 800 and 1000 nm, as schematically apparent from the filter characteristic illustrated in Fig. 2.
- Such filters can be manufactured by i.a. Omega Optical, Inc., Brattleboro, VT, U.S.A..
Abstract
Description
Claims
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP95941309A EP0805979A1 (en) | 1994-12-08 | 1995-12-08 | Arrangement in a gel electrophoresis apparatus |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
SE9404274A SE9404274D0 (en) | 1994-12-08 | 1994-12-08 | Device at a gel electrophoresis apparatus |
SE9404274-4 | 1994-12-08 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1996018099A1 true WO1996018099A1 (en) | 1996-06-13 |
Family
ID=20396268
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/SE1995/001482 WO1996018099A1 (en) | 1994-12-08 | 1995-12-08 | Arrangement in a gel electrophoresis apparatus |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP0805979A1 (en) |
SE (1) | SE9404274D0 (en) |
WO (1) | WO1996018099A1 (en) |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0840115A2 (en) * | 1996-11-05 | 1998-05-06 | Hitachi Electronics Engineering Co., Ltd. | DNA base sequencer |
WO1998023950A1 (en) * | 1996-11-29 | 1998-06-04 | Oxford Glycosciences (Uk) Ltd. | Gels, methods and apparatus for identification and characterization of biomolecules |
US6438259B1 (en) * | 1997-06-24 | 2002-08-20 | Large Scale Biology Corporation | Computer-assisted methods and apparatus for identification and characterization of biomolecules |
US6459994B1 (en) | 1998-05-29 | 2002-10-01 | Oxford Glycosciences (Uk) Ltd | Methods for computer-assisted isolation of proteins |
US9295393B2 (en) | 2012-11-09 | 2016-03-29 | Elwha Llc | Embolism deflector |
US10531655B2 (en) | 2011-12-02 | 2020-01-14 | The Regents Of The University Of California | Reperfusion protection solution and uses thereof |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1987007719A1 (en) * | 1986-06-03 | 1987-12-17 | Europäisches Laboratorium Für Molekularbiologie (E | Apparatus for the detection of substances which can be excited for photon emission |
EP0275440A2 (en) * | 1987-01-23 | 1988-07-27 | Hitachi, Ltd. | Photo detection system for DNA base analysis |
US5062942A (en) * | 1989-04-12 | 1991-11-05 | Hitachi, Ltd. | Fluorescence detection type electrophoresis apparatus |
EP0539743A1 (en) * | 1991-09-30 | 1993-05-05 | Beckman Instruments, Inc. | Enhanced fluorescence detection of samples in capillary column |
-
1994
- 1994-12-08 SE SE9404274A patent/SE9404274D0/en unknown
-
1995
- 1995-12-08 WO PCT/SE1995/001482 patent/WO1996018099A1/en not_active Application Discontinuation
- 1995-12-08 EP EP95941309A patent/EP0805979A1/en not_active Withdrawn
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1987007719A1 (en) * | 1986-06-03 | 1987-12-17 | Europäisches Laboratorium Für Molekularbiologie (E | Apparatus for the detection of substances which can be excited for photon emission |
EP0275440A2 (en) * | 1987-01-23 | 1988-07-27 | Hitachi, Ltd. | Photo detection system for DNA base analysis |
US5062942A (en) * | 1989-04-12 | 1991-11-05 | Hitachi, Ltd. | Fluorescence detection type electrophoresis apparatus |
EP0539743A1 (en) * | 1991-09-30 | 1993-05-05 | Beckman Instruments, Inc. | Enhanced fluorescence detection of samples in capillary column |
Non-Patent Citations (1)
Title |
---|
ANAL. CHEM., Volume 61, 1989, PATRICE L. CHRISTENSEN et al., "Fluorescence-Detected Circular Dichroism for On-Column Detection in Capillary Electrophoresis", pages 1344-1347. * |
Cited By (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0840115A3 (en) * | 1996-11-05 | 1999-08-25 | Hitachi Electronics Engineering Co., Ltd. | DNA base sequencer |
EP0840115A2 (en) * | 1996-11-05 | 1998-05-06 | Hitachi Electronics Engineering Co., Ltd. | DNA base sequencer |
US6480618B1 (en) | 1996-11-29 | 2002-11-12 | Oxford Glycosciences (Uk) Ltd. | Robotic device for removing selected portions of a polyacrylamide gel |
WO1998023950A1 (en) * | 1996-11-29 | 1998-06-04 | Oxford Glycosciences (Uk) Ltd. | Gels, methods and apparatus for identification and characterization of biomolecules |
US6064754A (en) * | 1996-11-29 | 2000-05-16 | Oxford Glycosciences (Uk) Ltd. | Computer-assisted methods and apparatus for identification and characterization of biomolecules in a biological sample |
US6278794B1 (en) | 1996-11-29 | 2001-08-21 | Oxford Glycosciences (Uk) Ltd | Computer-assisted isolation and characterization of proteins |
US6643391B2 (en) * | 1997-06-24 | 2003-11-04 | Large Scale Proteomics Corporation | Apparatus for computer-assisted isolation and characterization of proteins |
US6507664B1 (en) * | 1997-06-24 | 2003-01-14 | Large Scale Biology Corporation | Two-dimensional gels for separation, identification and characterization of biomolecules |
US6438259B1 (en) * | 1997-06-24 | 2002-08-20 | Large Scale Biology Corporation | Computer-assisted methods and apparatus for identification and characterization of biomolecules |
US6459994B1 (en) | 1998-05-29 | 2002-10-01 | Oxford Glycosciences (Uk) Ltd | Methods for computer-assisted isolation of proteins |
US10531655B2 (en) | 2011-12-02 | 2020-01-14 | The Regents Of The University Of California | Reperfusion protection solution and uses thereof |
US9295393B2 (en) | 2012-11-09 | 2016-03-29 | Elwha Llc | Embolism deflector |
US9414752B2 (en) | 2012-11-09 | 2016-08-16 | Elwha Llc | Embolism deflector |
Also Published As
Publication number | Publication date |
---|---|
EP0805979A1 (en) | 1997-11-12 |
SE9404274D0 (en) | 1994-12-08 |
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