Rapid and automatic fluorescence immunoassay analyzer for multiple micro-samples

• US 4,626,684 A
• Filed: 07/13/1983
• Issued: 12/02/1986
• Est. Priority Date: 07/13/1983
• Status: Expired due to Fees

• Current Assignee: Flow Laboratories Inc. (Bausch Health Cos., Inc.)
• Original Assignee: Isaac J. Landa
• Inventors: Landa, Isaac J.
• Primary Examiner(s): FALCONE, JANICE ARRINGTON
• Application Number:
  US06/513,307
• Time in Patent Office: 1,238 Days
• Field of Search: 356/317, 356/318, 250/461.2, 250/372, 250/328, 250/458.1, 250/461.1, 350/96.18
• US Class Current: 250/328
• CPC Class Codes: G01J 3/1833 Grazing incidence
• CPC Class Codes: G01J 3/4406 Fluorescence spectrometry
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 2021/6476 Front end, i.e. backscatter, geometry
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 2201/0446 Multicell plate, sequential

Abstract

An apparatus uses a rapid and efficient method for conducting fluorescence measurements, such as immuno assay, of a plurality of samples of a substance using pulsed or continuous light energy, front-end fluorescent optics, a mechanism which allows for lateral and tranverse scanning of rows and columns of wells, and spectra emission analysis of the spectra from the excited samples.

Claims

1. A fluorescence measuring apparatus for identifying a substance including:

excitation means for producing light energy;

carrier means for holding a plurality of samples which include said substance during excitation scanning of said samples;

front-end fluorescent optics for directing said excitation light energy on a path orthogonal to the surface of said substance and for receiving fluorescent emission spectra back along substantially the same path from said substance;

means for scanning said front-end fluorescent optics from sample to sample of said plurality of said samples which include said substance; and

spectra emission analyzing means for receiving fluorescent emission spectra from said excited samples which include said substance through said front-end fluorescent optics, said emission analyzing means having means for detecting light intensity at the measured wavelength of said fluorescent emission spectra.

2. The fluorescence measuring apparatus of claim 1 wherein said excitation means is a pulsed xenon lamp.

3. The fluorescence measuring apparatus of claim 1 wherein said carrier means is a plate having a plurality of wells adapted for holding samples which include said substance.

4. The fluorescence measuring apparatus of claim 3 wherein said scanning means causes said front-end fluorescent optics to move orthogonal to the direction of said movement of said carrier means while scanning each row of said plate.

5. The fluorescence measuring apparatus of claim 1 wherein said front-end fluorescent optics includes a fiber optics bundle comprised of source light conductors and detection light conductors.

6. The fluorescence measuring apparatus of claim 5 wherein said fiber optics bundle includes a central light source conductor surrounded by a plurality of detector light conductors.

7. The fluorescence measuring apparatus of claim 5 further including a lens for focusing the exiting and entering light from and to said fiber optics bundle.

8. The fluorescence measuring apparatus of claim 1 wherein said excitation means is a light source monochromator and said emission means is a detection monochromator.

9. The fluorescence measuring apparatus of claim 8 wherein said detection monochromator is tunable continuously over the operating spectral range.

10. The fluorescence measuring apparatus of claim 8 wherein said excitation monochromator is tunable continuously over the operating spectral range.

11. The fluorescence measuring apparatus of claim 8 wherein the light source monochromator includes exit and entrance slits the width of which are adjustable thereby providing adjustable wavelength bandpass.

12. The fluorescence measuring apparatus of claim 8 wherein the detection monochromator includes exit and entrance slits the width of which are adjustable thereby providing adjustable wavelength bandpass.

13. The fluorescence measuring apparatus of claim 8 wherein emission analyzing means further includes an electron photomultiplier tube.

14. The fluorescence measuring apparatus of claim 8 wherein said emission analyzing means further includes a solid state detector.

15. The fluorescence measuring apparatus of claim 1 further including reference detector means for compensating light intensity variations of said excitation means.

16. The fluorescence measuring apparatus of claim 1 wherein said scanning means transport said front-end fluorescent optics to any one of said plurality of samples held by said carrier means.

17. The fluorescence measuring apparatus of claim 1 wherein said scanner means can be adjusted to vary the measurement time for each sample.

18. The fluorescence measuring apparatus of claim 1 wherein said spectra emission analyzing means includes a microprocessor and a high speed sampling analyzer, said microprocessor being adapted to receive signals indicative of received fluorescent emission spectra from said high speed sampling analyzer.

19. The fluorescence measuring apparatus of claim 18 wherein said microprocessor processes said signals to subtract background noise and stray light effects from the measurements made for each sample.

20. The fluorescence measuring apparatus of claim 1 further including auto blank subtract means applied to the light intensity detecting means for subtracting background fluorescense from the sample fluorescence.

21. A fluorescence measuring apparatus for identifying a substance including:

excitation means for producing light energy;

carrier means for holding a plurality of samples which include said substance during excitation scanning of said samples;

front-end fluorescent optics for directing said excitation light energy on a path orthogonal to the surface of said substance and for receiving fluorescent emission spectra back along substantially the same path from said substance;

means for scanning said front-end fluorescent optics from sample to sample of said plurality of said samples which include said substance; and

fluorescence and phosphorescence life time measuring means for receiving fluorescent emissions from said excited samples which include said substance through said front-end fluorescent optics to determine the life time of said fluorescent phenomena.

22. Apparatus for fluorometric analysis of samples containing a substance comprising:

means for emitting a tunable, substantially monochromatic beam of radiation;

first fiber optic means for conducting said beam to a container of a sample to be analyzed;

lens means on one end of said fiber optics means for focusing said beam on the surface of said sample in said container;

second fiber optics means for detecting fluorescent radiation emitted by said substance;

said second fiber optics means being aligned with said first fiber optics means such that the radiation incident on said sample is substantially parallel to the emitted radiation detected by said second fiber optic means; and

means optically connected to said second fiber optic means for measuring the intensity of particular wavelength components of the emitted fluorescent radiation.