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
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
  • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
  • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
  • G01N21/64—Fluorescence; Phosphorescence
  • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"

Definitions

• the present invention relates to a method for detecting substances which are labelled with a fluorophore having a light absorption maximum at a wavelength between 480 and 520 nm by excitation of the fluorophore by means of light from a laser, and detection of light emitted by the fluorophore; an apparatus for detecting substances which are labelled with a fluorophore having a light absorption maximum at a wavelength between 480 and 520 nm, comprising a laser for excitation of the fluorophore by means of laser light, and a photodetector for detection of light emitted by the flurophore; and the use of a specific type of flurophores for labelling substances which are to be detected, together with a specific type of lasers for excitation of the fluorophores.
• argon lasers are bulky, expensive and power demanding and, therefore, are not suitable to be built into commercial electrophoresis apparatuses.
• Argon lasers have another disadvantage in that their output power is not stable but fluctuates. Moreover, argon lasers can not be pulsed which can be desirable in certain applications, e.g. time resolved fluorescence measurement.
• the object of the invention is therefore to define a laser for excitation of a fluorophore of the type mentioned in the introduction, which laser does not have the disadvantages mentioned above and which therefore is suitable to be built into commercial electrophoresis apparatuses.
• the object according to the invention is also attained by means of the apparatus according to the invention in that the laser for the excitation of the fluorophore, is either a Nd.YAG-laser emitting a a wavelength of 473 nm or a frequency-doubled laser diode emitting at a wavelength of
• the use according to the invention consists of the use of a fluorophore having a light absorption maximum at a wavelength between 480 and 520 nm for labelling substances to be detected, and either a Nd.YAG-laser emitting at a wavelength of 473 nm or a frequency-doubled laser diode emitting at a wavelength of 490 nm for excitation of the fluorophore.
• FIG. 1 schematically shows an embodiment of an apparatus according to the invention.
• 1 generally denotes a laser which emits a laser beam 2, which via a mirror 3 is directed into a gel 4 through one of its lateral edges, which gel 4 is provided between two glass plates 5 and 6.
• the gel 4 is included in an electrophoretic separation system, not shown in any greater detail, for separating substances which, according to the invention, are labelled with a fluorophore which has a light absorption maximum at a wavelength between 480 and 520 nm.
• Fluorescein isothiocyanate (FITC) , BODIPY-FL, YOYO-1 and TOTO-1, all available from the US company Molecular Probes, Inc., Oregon, U.S.A., are examples of such fluorophores.
• Either a Nd.YAG-laser emitting at a wavelength of 473 nm or a frequency-doubled laser diode emitting at a wavelength of 490 nm is preferably used as laser 1 according to the invention, both of which being well suited, due to their size, to be built into commercial gel electrophoresis apparatuses, particularly for separating nucleic acid fragments. These lasers emit blue light with small fluctuations and can, moreover, be pulsed.
• Another advantage of using lasers emitting blue light in comparison to lasers emitting green, yellow or red light, is that a better geometric resolution is obtained in the applications in question in that a blue laser beam is narrower than a red as well as a yellow and a green laser beam.
• fluorophore-laser combination defined in accordance with the invention, a combination is obtained which, what concerns size and efficiency, is extremely well suited for use in commercial electrophoresis apparatuses, particularly for the separation of nucleic acid fragments.

Priority Applications (2)

Applications Claiming Priority (2)

Publications (1)

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ID=20397051

Family Applications (1)

Country Status (4)

Cited By (2)

Citations (2)

• 1995
  • 1995-02-01 SE SE9500361A patent/SE9500361D0/sv unknown
• 1996
  • 1996-01-31 JP JP8523474A patent/JPH10513555A/ja active Pending
  • 1996-01-31 EP EP96902044A patent/EP0807247A1/en not_active Withdrawn
  • 1996-01-31 WO PCT/SE1996/000109 patent/WO1996024042A1/en not_active Application Discontinuation

Patent Citations (2)

Non-Patent Citations (1)

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