Interlaced lasers for multiple fluorescence measurement

• US 6,139,800 A
• Filed: 06/22/1998
• Issued: 10/31/2000
• Est. Priority Date: 06/23/1997
• Status: Expired due to Fees

• Current Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Original Assignee: Luminex Corporation (DiaSorin S.p.A.)
• Inventors: Chandler, Van S.
• Primary Examiner(s): SNAY, JEFFREY R
• Application Number:
  US09/102,034
• Time in Patent Office: 862 Days
• Field of Search: 422/82.08, 436/63, 436/172, 356/72, 356/73
• US Class Current: 422/82.08
• CPC Class Codes: G01N 15/1456 without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1477 Multiparameters
• CPC Class Codes: G01N 2015/1493 Particle size
• CPC Class Codes: G01N 2021/4707 Forward scatter; Low angle scatter
• CPC Class Codes: G01N 2021/4726 Detecting scatter at 90°
• CPC Class Codes: G01N 2021/6441 with two or more labels
• CPC Class Codes: G01N 2021/6482 Sample cells, cuvettes
• CPC Class Codes: G01N 21/53 within a flowing fluid, e.g. smoke
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters

Abstract

An apparatus for detecting at least one receptor in a biological sample in a flow stream for immunoassays, complex genetic analyses, and enzymatic assays. The biological sample includes a carrier having an indicator, such as a fluorescent dye, and a reactant that reacts to the receptor. The indicator indicates a presence of the receptor upon excitation by light having a wavelength or a band of wavelengths. The apparatus includes a viewing chamber for receiving the biological sample in the flow stream and for displaying the same. Light sources are arranged around the viewing chamber so as to emit light sequentially, simultaneously, or substantially simultaneously toward substantially identical or overlapping focal regions in the viewing chamber. At least one optical detector is configured to detect light emitted from the sample responsive to light emitted from the light sources. The apparatus includes band-pass filters for filtering out the emission wavelength reflecting the presence of the receptor and at least one multi-mode fiber optic cable connecting the optical detector to the filters.

Claims

1. An apparatus for detecting at least one receptor in at least one biological sample in a flow stream for at least one of immunoassays, complex genetic analyses, and enzymatic assays, the biological sample including at least one indicator indicating a presence of the at least one receptor upon excitation by at least one of a wavelength and band of wavelengths, said apparatus comprising:

a viewing chamber receiving the at least one biological sample and providing a plurality of focal regions therefor;

a plurality of light sources configured to emit light, one of sequentially, simultaneously, and substantially simultaneously, toward a substantially identical focal region of said plurality of focal regions in said viewing chamber on said at least one biological sample; and

a plurality of optical detectors in optical relationship with said plurality of light sources, respectively, and detecting a plurality of sample light emitted from said at least one sample responsive to the light emitted from the substantially identical focal region of said at least one biological sample.

2. The apparatus according to claim 1, further comprising:

a plurality of filter sets, each filter set including at least one band-pass filter connected to a respective optical detector, respectively; and

a plurality of multi-mode cables connecting said plurality of filter sets to said plurality of optical detectors, respectively.

3. The apparatus according to claim 2, wherein each band-pass filter is coupled to a respective amplifying photo-detector.

4. The apparatus according to claim 3, wherein each amplifying photo-detector includes one of a photo-multiplier tube and an avalanche photo-diode.

5. The apparatus according to claim 3, further comprising:

a plurality of analog-to-digital converters, said each amplifying photo-detector connected to a respective analog-to-digital converter;

a digital signal processor connected to said plurality of analog-to-digital converters; and

a selector connected to said plurality of analog-to-digital converters and to said digital signal processor.

6. The apparatus according to claim 1, wherein said plurality of light sources includes one of an optical amplifier and an arc lamp.

7. The apparatus according to claim 6, wherein said plurality of optical amplifiers includes at least one of a laser diode and a gas laser.

8. The apparatus according to claim 2, further comprising:

a plurality of multiplexers, each multiplexer including inputs, connected to each band-pass filter of a respective filter set, and outputs;

a plurality of amplifying photo-detectors connected to the outputs of said plurality of multiplexers;

a plurality of analog-to-digital converters connected to said plurality of amplifying photo-detectors;

a digital signal processor connected to said plurality of analog-to-digital converters; and

a selector connected to said digital signal processor and to said plurality of analog-to-digital converters.

9. The apparatus according to claim 2, wherein said plurality of multi-mode cables includes a multi-mode fiber optic cable.

10. The apparatus according to claim 1, wherein said viewing chamber comprises a quartz cuvette.

11. The apparatus according to claim 1, wherein said plurality of light sources and said plurality of optical detectors are substantially circumferentially and substantially perpendicularly arranged relative to a flow direction in said viewing chamber.

12. An apparatus for detecting at least one receptor in at least one biological sample in a flow stream for at least one of immunoassays, complex genetic analyses, and enzymatic assays, the at least one biological sample including at least one indicator indicating a presence of the at least one receptor upon excitation by a wavelength or band of wavelengths, said apparatus comprising:

a viewing chamber receiving the at least one biological sample in the flow stream;

a plurality of light sources, each directed to one of substantially identical and overlapping focal region with respect to said viewing chamber relative to remaining light sources of said plurality of light sources; and

at least one optical detector detecting light at least one of emitted and reflected from the at least one biological sample.

13. The apparatus according to claim 12, further comprising:

a plurality of band-pass filters operatively connected to said at least one optical detector; and

a multi-mode fiber optic cable including a number of bundles at least equal to a number of said plurality of band-pass filters, said multi-mode fiber optic cable connecting said optical detector to said plurality of band-pass filters.

14. The apparatus according to claim 13, further comprising:

a multiplexer including inputs connected to said plurality of band-pass filters;

a photo-detector, said multiplexer including an output connected to said photo-detector;

an analog-to-digital converter connected to said photo-detector; and

a digital signal processor connected to said analog-to-digital converter.

15. The apparatus according to claim 13, further comprising:

a plurality of photo-detectors, each photo-detector connected to a respective band-pass filter;

a plurality of analog-to-digital converters, each analog-to-digital converter connected to a respective photo-detector;

a digital signal processor connected to said plurality of analog-to-digital converters; and

a selector connected to said plurality of analog-to-digital converters and to said digital signal processor.

16. An apparatus for detecting at least two receptors in a biological sample transported in a flow stream for at least one of immunoassays, complex genetic analyses, and enzymatic assays, the biological sample including at least two indicators indicating a presence of the at least two receptors upon excitation by a wavelength or band of wavelengths, said apparatus comprising:

a viewing chamber receiving the biological sample transported via the flow stream;

a plurality of light sources arranged to emit, one of sequentially, simultaneously, and substantially simultaneously, light having at least two focal regions on the biological sample so as to illuminate the biological sample at a same or substantially same location in said viewing chamber, said plurality of light sources having wavelengths or bands of wavelengths selected to excite the at least two indicators in the biological sample by emitting the light on the two focal regions;

at least one optical detector optically responsive to said plurality of light sources so as to detect the presence of the at least two receptors.

17. The apparatus according to claim 16, further comprising:

a plurality of filter sets, each filter set including at least one band-pass filter connected to a respective optical detector, respectively; and

a plurality of multi-mode cables connecting said plurality of filter sets to said plurality of optical detectors, respectively.

18. The apparatus according to claim 17, further comprising:

a plurality of amplifying photo-detectors, each amplifying photo-detector operatively coupled to a respective band-pass filter;

a plurality of analog-to-digital converters, said each amplifying photo-detector connected to a respective analog-to-digital converter;

a digital signal processor connected to said plurality of analog-to-digital converters; and

a selector connected to said plurality of analog-to-digital converters and to said digital signal processor.

19. The apparatus according to claim 17, further comprising:

a plurality of multiplexers, each multiplexer including inputs, connected to each band-pass filter of a respective filter set, and outputs;

a plurality of amplifying photo-detectors connected to the outputs of said plurality of multiplexers;

a plurality of analog-to-digital converters connected to said plurality of amplifying photo-detectors;

a digital signal processor connected to said plurality of analog-to-digital converters; and

a selector connected to said digital signal processor and to said plurality of analog-to-digital converters.

20. The apparatus according to claim 17 wherein the at least two focal regions are one of non-overlapping and overlapping.

21. An apparatus for detecting at least two receptors in at least one biological sample in a flow stream for at least one of immunoassays, complex genetic analyses, and enzymatic assays, the at least one biological sample including at least two indicators indicating a presence of the at least two receptors upon excitation, said apparatus comprising:

at least one light source emitting at least two light rays substantially simultaneously toward at least two regions on said at least one biological sample;

at least one optical detector detecting at least two light rays one of sequentially, simultaneously, and substantially simultaneously, emmitted from the at least two regions on the at least one biological sample via detecting the at least two indicators indicating the presence of the at least two receptors for the at least one of immunoassays, complex genetic analyses, and enzymatic assays.

22. An apparatus for detecting at least two receptors in at least one biological sample in a flow stream for at least one of immunoassays, complex genetic analyses, and enzymatic assays, the at least one biological sample including at least two indicators indicating a presence of the at least two receptors upon excitation, said apparatus comprising:

first means for emitting at least two light rays one of sequentially, simultaneously, and substantially simultaneously, toward a substantially identical region on said at least one biological sample;

second means for detecting at least two light rays one of sequentially, simultaneously, and substantially simultaneously, emmitted from the substantially identical region on the at least one biological sample via detecting the at least two . indicators indicating the presence of the at least two receptors for the at least one of immunoassays, complex genetic analyses, and enzymatic assays.