Electrophoresis method and apparatus having continuous detection means

• US 4,707,235 A
• Filed: 10/16/1985
• Issued: 11/17/1987
• Est. Priority Date: 10/16/1985
• Status: Expired due to Term

• Current Assignee: Pharmacia Limited (Pfizer Inc.)
• Original Assignee: Pharmacia Limited (Pfizer Inc.)
• Inventors: Englert, David F., Wheeler, Richard J.
• Primary Examiner(s): Tung, T.
• Application Number:
  US06/787,904
• Time in Patent Office: 762 Days
• Field of Search: 204/299 R, 204/182.8, 204/182.7, 250/374, 250/375
• US Class Current: 204/461
• CPC Class Codes: G01N 27/44717 Arrangements for investigating the separated zones, e.g. localising zones
• CPC Class Codes: G01N 27/44782 of a plurality of samples

Abstract

A multi-channel electrophoresis apparatus is fitted with a detection means disposed substantially across the width of the medium, intersecting all channels. The detection means senses components of samples as they migrate past the detection means and identifies the channel in which the detected components are located. In the preferred embodiment, the samples are tagged with radioisotopes which are detected by the detector.

Claims

1. An apparatus for analytical electrophoresis comprising:

a layer of electrophoresis medium having a plurality of channels, each of which is adapted to electrophoretically separate samples into components along said channels in the plane of said layer;

means to introduce a plurality of samples having multiple components into one end of said medium in said plurality of channels;

means to apply an electric potential to said medium along said channels;

detection means disposed in a fixed position substantially across the width of said medium, intersecting said channels, said detection means being substantially separated from said sample introduction means, said detection means adapted to detect continuously in time during the electrophoretic migration said components of said samples in said channels and to identify the channel in which the detected components are located as said components pass through said medium adjacent to said detection means.

2. The apparatus as described in claim 1, wherein the components of said samples are labeled.

3. The apparatus as described in claim 2, wherein the components of said samples are radioactively labeled.

4. The apparatus as described in claim 3, wherein the detector comprises an integral position-sensitive radioisotope detector.

5. The apparatus as described in claim 4, wherein said detector comprises a narrow, elongated chamber for gas disposed within one millimeter of the electrophoresis medium, ionizable gas within said chamber, means to electronically amplify the ionizations produced in the gas by ionizing radiation, and means for electronically identifying the channel position of the amplified electrical discharges along the length of the contained gas.

6. The apparatus as described in claim 5, wherein ionizing radiation is admitted to the chamber through an elongated slit in a radiation absorbing material.

7. The apparatus as described in claim 6, wherein said slit is about 0.5-2.0 mm wide.

8. The apparatus of claim 7, wherein the radiation absorbing material is lead.

9. The apparatus of claim 1, wherein multiple detectors are disposed across said medium.

10. The apparatus of claim 1, wherein said medium is homogeneous in composition along said channels.

11. The apparatus of claim 1, wherein the medium has a composition gradient along the length of said channels.

12. A method of continuously analytically observing the separation of samples into a plurality of components by electrophoresis simultaneously for a plurality of samples which comprises:

introducing samples containing multiple components into separate channels in an electrophoresis medium;

subjecting said samples to an electric potential in said channels of said electrophoresis medium to cause components of each sample to migrate along said channel; and

continuously in time detecting said components in each sample simultaneously during the electrophoretic migration, at a location separated from the position of sample introduction with a detector disposed in a fixed position substantially across the width of said medium, as said components pass through said medium in their respective channels.

13. The method as described in claim 12, wherein the components of said samples are labeled.

14. The method as described in claim 13, wherein the components of said samples are radioactively labeled.

15. The method as described in claim 14, wherein said components are labeled with phosphorus-32.

16. The method as described in claim 14 or 15, wherein passage of said labeled components of all samples are detected with a position-sensitive radioisotope detector disposed across said channels.

17. The method as described in claim 16, wherein the data collected by said detector is stored electronically.

18. The method as described in claim 17, wherein the data collected by said detector is accumulated for 30 seconds prior to storage.

19. The method as described in claim 18, wherein said collected data is displayed as a function of time.

20. The method as described in claim 12, wherein the electrical potential is maintained at the same level throughout the separation.

21. The method as described in claim 12, wherein the temperature of the electrophoresis medium is maintained at the same temperature throughout the separation.

22. Method of sequencing nucleic acids which comprises:

introducing samples containing nucleotide chains into separate channels in an electrophoresis medium;

subjecting said samples to an electric potential in said channels of said electrophoresis medium to cause nucleotide chains of each sample to migrate along said channel; and

continuously in time detecting said nucleotide chains in each channel simultaneously during the electrophoretic migration, at a location separated from the position of sample introduction with a detector disposed in a fixed position substantially across the width of said medium, as said nucleotide chains pass through said medium in their respective channels.

23. The method as described in claim 22, wherein said nucleotide chains are labeled.

24. The method as described in claim 23, wherein said nucleotide chains are radioactively labeled.

25. The method as described in claim 24, wherein said nucleotide chains are labeled with phosphorus-32.