Apparatus for gel electrophoresis

• US 5,294,323 A
• Filed: 06/01/1993
• Issued: 03/15/1994
• Est. Priority Date: 05/26/1993
• Status: Expired due to Fees

• Current Assignee: Hitachi High-Tech Electronics Engineering Co., Ltd. (Hitachi, Ltd.)
• Original Assignee: Hitachi High-Tech Electronics Engineering Co., Ltd. (Hitachi, Ltd.), Hitachi, Ltd.
• Inventors: Togusari, Teruo, Irisawa, Kazuyuki, Kambara, Hideki
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US08/069,841
• Time in Patent Office: 287 Days
• Field of Search: 204/299 R, 204/182.8, 356/344
• US Class Current: 204/612
• CPC Class Codes: G01N 27/44721 by optical means

Abstract

In an apparatus for gel electrophoresis in which a sample of fluorophore-labelled DNA fragments is caused to migrate by electrophoresis through a gel electrolyte layer in an electrophoresis plate from top to bottom, thereby separating the sample into individual DNA fragments, and a laser beam is launched horizontally into said electrolyte layer from one side of the electrophoresis plate in a direction perpendicular to the longitudinal axis of said electrophoresis plate, with the emitted fluorescences being detected to determine the base sequences of the respective DNAS, a mirror for reflecting fluorescences is provided at the back of the electrophoresis plate in such a way that it is parallel to the direction of laser beam application, and the fluorescences reflected by this mirror are received by a fluorescence detector such as a CCD sensor or MOS linear image sensor. In one embodiment, the mirror is inclined by an angle of 45° with the electrophoresis plate in such a way that it reflects the incident fluorescences upward, and the fluorescence detector is positioned above the mirror along the electrophoresis plate. In another embodiment, Texas Red having an excitation maximum wavelength of 596 nm and an emission maximum wavelength of 615 nm is used as the fluorophore and a He-Ne laser operating at a wavelength of 594 nm is used as the light source.

Claims

1. In an apparatus for gel electrophoresis in which a sample of fluorophore-labelled DNA fragments is caused to migrate by electrophoresis through a gel electrolyte layer in an electrophoresis plate in a first direction, thereby separating the sample into individual DNA fragments, and laser light is launched into a linear irradiation region of said gel electrolyte layer in a second direction which is generally perpendicular to said first direction with the emitted fluorescence being detected to determine the base sequence of the particular DNA, the improvement wherein a mirror for reflecting fluorescence is provided at the back of the electrophoresis plate in such a way that it is parallel to said linear irradiation region and the fluorescence reflected from said mirror is received by a separate fluorescence detector composed of a solid-state imaging device.

2. An apparatus for gel electrophoresis according to claim 1 wherein said fluorophore is Texas Red having an excitation maximum wavelength of 596 nm and an emission maximum wavelength of 615 nm, and a He-Ne laser operating at a wavelength of 594 nm is used as a laser light source.

3. An apparatus for gel electrophoresis according to claim 1 wherein said mirror is inclined by an angle of about 45°

with the electrophoresis plate so that it reflects the incident fluorescence upward, and said fluorescence detector is positioned above said mirror along the electrophoresis plate.

4. An apparatus for gel electrophoresis according to claim 1 wherein said mirror has the necessary and sufficient length to insure that the fluorescences from a predetermined number of pixels that are arranged to span opposite ends of the linear irradiation region of the electrophoresis plate will be reflected upward in one action.

5. An apparatus for gel electrophoresis according to claim 1 wherein said solid-state imaging device is a CCD sensor or a MOS linear image sensor and is cooled with a mechanism composed of a Peltier device.

6. An apparatus for gel electrophoresis according to claim 1 wherein signal processing boards are provided adjacent said mirror and said fluorescence detector.

7. An apparatus for gel electrophoresis according to claim 1 wherein DNA fragments of a concentration within the range from 0.1 to 0.15 picomole and fluorophore of a concentration within the range from 0.1 to 0.15 picomole are injected into grooves associated with individual electrophoresis lanes.