Detection apparatus for electrophoretic gels

• US 5,162,654 A
• Filed: 02/01/1991
• Issued: 11/10/1992
• Est. Priority Date: 02/01/1991
• Status: Expired due to Term

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Kostichka, Anthony J., Smith, Lloyd M.
• Primary Examiner(s): Fields, Carolyn E.
• Assistant Examiner(s): NOT, DEFINED
• Application Number:
  US07/650,393
• Time in Patent Office: 648 Days
• Field of Search: 250/461.2, 250/461.1, 250/458.1, 250/302, 250/303, 204/299 R, 204/182.8
• US Class Current: 250/458.1
• CPC Class Codes: G01J 2003/1226 Interference filters
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 2021/6441 with two or more labels
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 27/44721 by optical means

Abstract

An apparatus is disclosed to optically determine which of four fluorophores is attached to a band of DNA molecules on an electrophoresis gel. The apparatus includes four separate band pass interference filters and four wedge prisms to create four discrete areas of light on a detector. Digital comparison of the relative intensity of light sensed in the four discrete areas determines the identity of the excited fluorophore.

Claims

1. An instrument for the detection of the fluorescence pattern emitted by biological molecules on an electrophoresis gel which are tagged with one of four fluorophores comprising:

excitation means to excite the fluorophores on the molecules;

an interference filter assembly which includes four interference filters selected to pass light in a wavelength band corresponding to the emission characteristics of the fluorophores;

a prism wedge assembly which includes a respective prism wedge optically associated with each of the interference filters in the interference filter assembly to bend the light signal from the associated interference filter so that the light signals emitted by the four interference filters are separated from each other;

a light detector for detecting light incident thereon and for determining the relative intensity of the light incident thereon; and

optical focusing means to direct fluorescence from the gel through the interference filter assembly and then through the prism wedge assembly to the detector so that the light impinging on the detector is separated into four bands, each of which is related to the intensity of the fluorescence from the gel in one of the wavelength bands associated with one of the interference filters.

2. An instrument as claimed in claim 1 wherein the excitation means is a laser.

3. An instrument as claimed in claim 1 wherein the interference assembly is made up of four laterally adjacent interference filters.

4. An instrument as claimed in claim 3 wherein the interference filters pass light at wavelengths centered at 540, 560, 580 and 610 nanometers respectively.

5. An instrument as claimed in claim 1 wherein the light detector is a CCD camera.

6. An instrument as claimed in claim 1 further comprising a digital computer to collect the data from the detector, the computer programmed to make a digital comparison of light received at discrete locations on the detector.