Apparatus and method for detecting nucleic acid amplification products

• US 6,015,674 A
• Filed: 03/20/1998
• Issued: 01/18/2000
• Est. Priority Date: 04/29/1994
• Status: Expired due to Term

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: The Perkin-Elmer Corporation Applied Biosystems Division
• Inventors: Young, Eugene F., Tracy, David H., Woudenberg, Timothy M., Shigeura, John, Bodner, Kevin S., Connell, Charles R.
• Primary Examiner(s): Horlick, Kenneth R.
• Application Number:
  US09/046,114
• Time in Patent Office: 669 Days
• Field of Search: 435/6, 435/91.2, 422/82.05, 422/82.08, 422/82.09
• US Class Current: 435/6.12
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 1/6851 Quantitative amplification
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2537/143 Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
• CPC Class Codes: C12Q 2545/101 with an internal standard/control
• CPC Class Codes: C12Q 2561/101 Taqman
• CPC Class Codes: G01N 2021/6484 Optical fibres
• CPC Class Codes: G01N 21/0303 Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment G01N21/031 - G01N21/15 take precedence
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters

Abstract

An apparatus is provided which includes a sample holder for holding a reaction chamber which includes an optical interface, a fiber optic cable for delivering an excitation beam to a sample housed within the reaction chamber and for receiving light emitted by the sample, and a lens co-axially disposed with the fiber optic cable and positioned outside the reaction chamber for focusing the excitation beam through the optical interface and within a volume of the sample and for collecting and transmitting to the fiber optic cable light emitted within the volume of the sample.

Claims

1. An apparatus comprising:

a sample holder for holding a sealed reaction chamber, the sealed reaction chamber including a sample, an optical interface and an air gap between the sample and the optical interface;

a fiber optic cable positioned outside the sealed reaction chamber to deliver an excitation beam to the sample housed within the reaction chamber and to receive light emitted by the sample; and

a lens co-axially disposed with the fiber optic cable and positioned outside the reaction chamber to focus the excitation beam through the optical interface and the air gap into a volume of the sample and to collect and transmit to the fiber optic cable through the optical interface and the air gap light emitted within the volume of the sample.

2. The apparatus according to claim 1 wherein the lens is not in contact with the reaction chamber.

3. The apparatus according to claim 1 wherein the lens is separated from the reaction chamber by an air gap.

4. The apparatus according to claim 1 wherein the apparatus further includes a mechanism for heating the optical interface to reduce condensation of the sample on the optical interface.

5. The apparatus according to claim 1 wherein the sample holder includes a removable reaction chamber for holding the sample.

6. The apparatus according to claim 1 wherein the lens is positioned above the sample holder.

7. The apparatus according to claim 1 wherein the apparatus further includes a detection and analysis mechanism for receiving first and second fluorescent signals from the fiber optic cable.

8. An apparatus comprising:

a sealable reaction chamber which includes an optical interface;

a fiber optic cable positioned outside the sealable reaction chamber to deliver an excitation beam to a sample housed within the reaction chamber and to receive light emitted by the sample; and

a lens co-axially disposed with the fiber optic cable and positioned outside the reaction chamber to focus the excitation beam through the optical interface into a volume of the sample and to collect and transmit to the fiber optic cable through the optical interface light emitted within the volume of the sample.

9. The apparatus according to claim 8 wherein the lens is not in contact with the reaction chamber.

10. The apparatus according to claim 8 wherein the lens is separated from the reaction chamber by an air gap.

11. The apparatus according to claim 8 wherein the apparatus further includes a mechanism for heating the optical interface to reduce condensation of the sample on the optical interface.

12. The apparatus according to claim 8 wherein the optical interface is positioned on a top of the reaction chamber and the lens is positioned above the optical interface.

13. The apparatus according to claim 8 wherein the reaction chamber is removable from the apparatus.

14. The apparatus according to claim 8 wherein the apparatus further includes a detection and analysis mechanism for receiving first and second fluorescent signals from the fiber optic cable.

15. An apparatus comprising:

a plurality of sample holders for holding a plurality of sealed reaction chambers, each sealed reaction chamber including a sample, an optical interface and an air gap between the sample and the optical interface;

a plurality of fiber optic cables positioned outside the sealed reaction chambers to deliver an excitation beam to samples housed within the plurality of reaction chambers and to receive light emitted by the samples; and

a plurality of lenses co-axially disposed with the fiber optic cables and positioned outside the reaction chambers to focus the excitation beams through the optical interfaces and the air gaps into volumes of the samples and to collect and transmit to the fiber optic cables through the optical interface and the air gap light emitted within the volumes of the samples.

16. The apparatus according to claim 15 wherein the apparatus further includes a fiber optic multiplexer for coupling a detection and analysis mechanism to each of the plurality of fiber optic cables.

17. The apparatus according to claim 15 wherein the plurality of lenses are not in contact with the reaction chambers.

18. The apparatus according to claim 15 wherein the plurality of lenses are separated from the reaction chambers by air gaps.

19. The apparatus according to claim 15 wherein the plurality of lenses are positioned above the sample holders.

20. The apparatus according to claim 15 wherein the apparatus further includes a mechanism for heating the optical interface to reduce condensation of the sample on the optical interface.

21. The apparatus according to claim 15 wherein the plurality of sample holders include one or more removable reaction chambers for holding the sample.

22. The apparatus according to claim 15 wherein the apparatus further includes a detection and analysis mechanism for receiving first and second fluorescent signals from the fiber optic cable.

23. An apparatus comprising:

a plurality of sealable reaction chambers which each include an optical interface;

a plurality of fiber optic cables positioned outside the sealable reaction chambers to deliver excitation beams to samples housed within the plurality of reaction chambers and to receive light emitted by the samples; and

a plurality of lenses co-axially disposed with the fiber optic cables and positioned outside the reaction chambers to focus the excitation beams through the optical interfaces into volumes of samples housed within the plurality of reaction chambers and to collect and transmit to the fiber optic cables through the optical interfaces light emitted within the volumes of the samples.

24. The apparatus according to claim 23 wherein the lenses are not in contact with the reaction chambers.

25. The apparatus according to claim 23 wherein the lenses are separated from the reaction chambers by air gaps.

26. The apparatus according to claim 23 wherein the reaction chambers are removable from the apparatus.

27. The apparatus according to claim 23 wherein the lenses are positioned above the sample holders.

28. A method for monitoring the formation of a nucleic acid amplification reaction product comprising:

amplifying a nucleic acid sequence in a sample in a sealed reaction chamber which includes an optical interface and an air gap between the sample and the optical interface;

delivering an excitation beam from a fiber optic cable positioned outside the sealed reaction chamber into a volume of the sample through the optical interface and the air gap; and

collecting light emitted by the volume of sample within the fiber optic cable through the optical interface and the air gap.

29. The method according to claim 28 wherein delivering and collecting including passing the excitation beam and light through a lens which is positioned outside the sealed reaction chamber between the fiber optic cable and the reaction chamber.

30. The method according to claim 29 wherein the lens is not in contact with the reaction chamber.

31. The method according to claim 28 wherein delivering and collecting is performed above the reaction chamber.

32. The method according to claim 28 further including the step of heating the optical interface to reduce condensation of the sample on the optical interface.