Electrophoresis and fluorescence detection apparatus

• US 5,712,476 A
• Filed: 03/27/1996
• Issued: 01/27/1998
• Est. Priority Date: 05/30/1995
• Status: Expired due to Term

• Current Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Original Assignee: Visible Genetics, Inc. (Bayer AG)
• Inventors: Cassidy, James W., Renfrew, John A.
• Primary Examiner(s): LE, QUE TAN
• Application Number:
  US08/624,789
• Time in Patent Office: 671 Days
• Field of Search: 250/214 R, 250/214 LA, 250/214 RC, 250/333, 250/214 C, 422/82.08, 422/82.07, 356/344, 356/222, 356/221
• US Class Current: 250/214.00R
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 30/8603 with integration or differentiation

Abstract

An improved electrophoresis and fluorescence detection apparatus has an electromagnetic radiation sensor juxtaposed with a sensing region. The output signal from the electromagnetic radiation sensor is a current signal, and the current signal is converted to a voltage signal. The voltage signal is summed with a programmable offset generated by an inexpensive eight-bit D/A converter. The offset signal is selected to establish a lower starting point for the dynamic range of the A/D conversion, and is selected to null some or all of the background electromagnetic radiation level. The summed signal is amplified and integrated in an integrator. The integrator is switchable under program control. The integrator is switched on for long and short intervals. The short intervals permit sensing over a dynamic range accommodating very high levels of fluorescence; very high peaks may be measured and features of the peaks distinguished. The long intervals permit sensing over a dynamic range that is optimized for the peaks associated with the smaller peaks of individual nucleotides. In this way, the dynamic range of the A/D convertor is set to permit the highest possible resolution over the range of interest during the time in which the sequencing of the nucleotides takes place. The sequencer is fast and economical and yields data with high resolution.

Claims

1. A detection apparatus comprising:

an electromagnetic radiation emitter, an electromagnetic radiation detector, and a detection area juxtaposed with the emitter and juxtaposed with the detector, said electromagnetic radiation detector having an electrical output indicative of detected light; and

an integrator responsive to the output, said integrator integrating over repeated short intervals and repeated long intervals, the short intervals being less than half as long in duration as the long intervals.

2. The detection apparatus of claim 1 wherein the short intervals are less than one-third as long in duration as the long intervals.

3. The detection apparatus of claim 1 wherein the short sampling intervals and the long intervals alternate.

4. A detection apparatus comprising:

a plurality of electromagnetic radiation emitters, an electromagnetic radiation detector, and a plurality of detection areas corresponding in number to the electromagnetic radiation emitters, each detection area juxtaposed with the electromagnetic radiation detector and juxtaposed with a corresponding electromagnetic radiation emitter, said electromagnetic radiation detector having an electrical output indicative of detected electromagnetic radiation; and

an integrator responsive to the output, said integrator integrating over repeated short intervals and repeated long intervals, the short intervals being less than half as long in duration as the long intervals;

said apparatus further comprising an electromagnetic radiation source and electromagnetic radiation switches coupling the electromagnetic radiation source with the electromagnetic radiation emitters, said electromagnetic radiation switches disposed such that during each interval, only one of the electromagnetic radiation emitters is coupled with the electromagnetic radiation source.

5. The detection apparatus of claim 4 wherein the short intervals are less than one-third as long in duration as the long intervals.

6. The detection apparatus of claim 5 wherein the short intervals and the long intervals alternate.

7. A detection apparatus comprising:

an electromagnetic radiation emitter, an electromagnetic radiation detector, and a detection area juxtaposed with the emitter and juxtaposed with the detector, said electromagnetic radiation detector having an electrical output indicative of detected electromagnetic radiation;

a programmable offset source having an offset output;

a summer summing the detector output and the offset output and yielding a summer output; and

an analog-to-digital converter responsive to the summer output.

8. The detection apparatus of claim 7 wherein the programmable offset source comprises an eight-bit digital-to-analog converter.

9. The detection apparatus of claim 7 wherein the analog-to-digital convertor has a twelve-bit output.

10. The detection apparatus of claim 7 further comprising an integrator electrically coupled between the summer and the analog-to-digital converter.

11. A detection apparatus comprising:

a plurality of electromagnetic radiation emitters, an electromagnetic radiation detector, and a plurality of detection areas corresponding in number to the electromagnetic radiation emitters, each detection area juxtaposed with the electromagnetic radiation detector and juxtaposed with a corresponding electromagnetic radiation emitter, said electromagnetic radiation detector having an electrical output indicative of detected electromagnetic radiation;

a programmable offset source having an offset output, a summer summing the detector output and the offset output and yielding a summer output; and

an analog-to-digital converter responsive to the summer output;

said apparatus further comprising an electromagnetic radiation source and electromagnetic radiation switches coupling the electromagnetic radiation source with the electromagnetic radiation emitters, said electromagnetic radiation switches disposed such that during each interval, only one of the electromagnetic radiation emitters is coupled with the electromagnetic radiation source.

12. The detection apparatus of claim 11 wherein the programmable offset source comprises an eight-bit digital-to-analog converter.

13. The detection apparatus of claim 11 wherein the analog-to-digital convertor has a twelve-bit output.

14. The detection apparatus of claim 11 further comprising an integrator electrically coupled between the summer and the analog-to-digital converter.