Electronic method for autofluorography of macromolecules on two-D matrices

• US 4,389,670 A
• Filed: 12/30/1981
• Issued: 06/21/1983
• Est. Priority Date: 12/30/1981
• Status: Expired due to Term

• Current Assignee: United States Of America As Represented By The Department Of Energy
• Original Assignee: United States Department of Energy
• Inventors: Davidson, Jackson B., Case, Arthur L.
• Primary Examiner(s): BRITTON, HOWARD W
• Application Number:
  US06/335,998
• Time in Patent Office: 538 Days
• Field of Search: 358/111, 358/107, 358/211, 235/92 PC, 378/49
• US Class Current: 348/162
• CPC Class Codes: G01N 21/5907 Densitometers
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 30/95 Detectors specially adapted therefor; Signal analysis
• CPC Class Codes: G01T 1/2942 using autoradiographic methods

Abstract

A method for detecting, localizing, and quantifying macromolecules contained in a two-dimensional matrix is provided which employs a television-based position sensitive detection system. A molecule-containing matrix may be produced by conventional means to produce spots of light at the molecule locations which are detected by the television system. The matrix, such as a gel matrix, is exposed to an electronic camera system including an image-intensifier and secondary electron conduction camera capable of light integrating times of many minutes. A light image stored in the form of a charge image on the camera tube target is scanned by conventional television techniques, digitized, and stored in a digital memory. Intensity of any point on the image may be determined from the number at the memory address of the point. The entire image may be displayed on a television monitor for inspection and photographing or individual spots may be analyzed through selected readout of the memory locations. Compared to conventional film exposure methods, the exposure time may be reduced 100-1000 times.

Claims

1. An autofluorographic method of detecting and quantifying labeled macromolecules resolved in a matrix in which light spots are produced which correspond to the location and quantity of said macromolecules in said matrix, including the steps of:

exposing said matrix to a television camera system having a lensless, intensified, light sensitive target means on which an electric charge image is accumulated corresponding to said light patterns of said matrix;

scanning said target in a raster with an electron beam to remove the accumulated charge image from said target, and

producing output signals indicative of the location and quantity of the electric charge removed from said charge image on said target.

2. The method of claim 1 wherein the exposure step further includes exposing said camera to said light patterns of said matrix for a preselected period to integrate the light emitted from said matrix prior to said scanning step.

3. The method of claim 1 further including the steps of converting said output signals to digital signals and storing said digital signals in a digital memory.

4. The method of claim 1 wherein said matrix includes a gel sheet having a scintillating material incorporated therein and wherein said macromolecules are radioactivity labeled to produce scintillations in said matrix at the locations of said macromolecules in said gel sheet.

5. The method of claim 4 wherein said matrix is formed of a polyacrylamide gel sheet and said macromolecules are tritium labeled proteins.

6. The method of claim 1 wherein said macromolecules are labeled with a fluorescent strain and further including the step of directing a source of light on said matrix to excite fluorescence of said stained molecules prior to exposing said matrix to said camera.

7. The method of claim 1 wherein said macromolecules are labeled with a phosphorescent stain.

8. The method of claim 1 wherein said macromolecules are labeled by induced radioactivity from neutron bombardment.

9. The method of claim 1 wherein said macromolecules are labeled with a stain that becomes visible in white light, and further including the step of directing a source of white light on said matrix during exposure to said camera.