Fluorometer

• US RE34,782 E
• Filed: 10/12/1992
• Issued: 11/08/1994
• Est. Priority Date: 07/01/1985
• Status: Expired due to Term

• Current Assignee: Diatron Corporation
• Original Assignee: Diatron Corporation
• Inventors: Dandliker, Walter B., Barr, Howard S., Katzenstein, Henry S., Watson, Keith R.
• Primary Examiner(s): Hannaher, Constantine
• Application Number:
  US07/969,794
• Time in Patent Office: 757 Days
• Field of Search: 250/458.1, 250/461.2, 356/318
• US Class Current: 250/458.1
• CPC Class Codes: G01N 21/6408 with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 2201/1087 Focussed scan beam, e.g. laser
• CPC Class Codes: G02B 21/16 adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence

Abstract

A fluorometer for measuring a particular fluorescence emanating from a specimen and operating in accordance with the following method. Producing a burst of concentrated light energy and directing the concentrated light energy toward the specimen to produce a fluorescence from the specimen including the particular fluorescence. Preferably producing an image of the fluorescence. Detecting the fluorescence and producing a signal in accordance with the fluorescence. Controlling the passage of the image of the fluorescence for detecting within a particular time period so as to optimize the detection of the particular fluorescence. Timing the operation to sequence the detection of the fluorescence within the particular time period after the production of the burst of concentrated light energy. Scanning the fluorescence from the specimen for forming signals representative of the fluorescence from the specimen. Analyzing the signals to enhance the portion of the signal representing the particular fluorescence relative to the portion of the signal.

Claims

1. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurements of the particular fluorescence from the measurements of the background fluorescence.Iaddend., including,

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the burst of concentrated light energy for directing the burst of concentrated light energy toward the specimen to produce a fluorescence from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen for detecting .[.the fluorescence.]. .Iadd.such fluorescences .Iaddend.and for producing signals in accordance with such detection,

means coupled to the detecting means for controlling the detecting means within a particular time period.Iadd., after the burst of the concentrated light energy, .Iaddend.to optimize the detection of the particular fluorescence and to enhance the detection of the particular fluorescence relative to the .[.total.]. .Iadd.detection of the background .Iaddend.fluorescence, the beginning of the particular time period being defined by a first particular time after the burst of concentrated light energy and during the production of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.and the end of the particular time period being defined by a second particular time after the first particular time where the second particular time occurs during the production of the particular fluorescence .[.from the specimen as a result of the burst of the concentrated light energy.].,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of .[.the above.]. .Iadd.such .Iaddend.means to sequence the detection of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.during the particular time period .[.as a result of.]. .Iadd.after .Iaddend.the production of the burst of concentrated light energy,

means .[.coupled to the detection means for forming.]. .Iadd.responsive to the signals produced in accordance with the detection during the particular time period for distinguishing the particular fluorescence produced during the particular time period from the background fluorescence produced during the particular time period and for producing output .Iaddend.signals representative .Iadd.substantially only .Iaddend.of the particular fluorescence from the specimen .Iadd.during the particular time period.Iaddend., and

means responsive to the .[.detection of.]. .Iadd.output signals representing substantially only .Iaddend.the particular fluorescence from the specimen during the particular time period for producing an image of the particular fluorescence.

2. The fluorometer of claim 1 wherein the means for .[.forming signals.]. .Iadd.detecting the fluorescences .Iaddend.includes means for producing a scanning of the particular .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen during the particular time period at different positions on the specimen.

3. The fluorometer of claim 2 wherein the means for detecting includes a photosensitive array for detecting the fluorescence emanating from a plurality of spots on the specimen and the means for scanning includes an array control for scanning the photosensitive array to obtain a reproduction of the detected fluorescence of the different spots on the specimen.

4. The fluorometer of claim 2 wherein the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the fluorescence emanating from the specimen at the different positions.

5. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurements of the particular fluorescence from the measurements of the background fluorescence, the particular fluorescence having different characteristics than the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the concentrated light energy for directing the concentrated light energy toward the specimen to produce .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence from the specimen .Iaddend.for detecting .[.the fluorescence .]. .Iadd.such fluorescences .Iaddend.and for producing signals in accordance with .[.the fluorescence.]. .Iadd.such detection.Iaddend.,

means coupled to the detecting means for controlling the detecting means within a particular time period .[.to optimize the detection of the particular fluorescence.]. .Iadd.after the burst of the concentrated light energy .Iaddend.and wherein the controlling means is an electro-optic modulator formed by a cubic crystal of the class T_d, .Iadd.the fluorescence produced during the particular time period including the particular fluorescence and the background fluorescence.Iaddend.,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the above means to sequence the detection of the fluorescence within the particular time period after the production of the burst of concentrated light energy, and

data processing means responsive to the signals produced by the detecting means for .[.analysing the signals to enhance.]. .Iadd.distinguishing .Iaddend.the portion of the signals .[.representing.]. .Iadd.produced during the particular time period in representation of .Iaddend.the particular fluorescence .[.relative to.]. .Iadd.from .Iaddend.the portion of the signals .[.representing the remaining.]. .Iadd.produced during the particular time period in representation of the background .Iaddend.fluorescence .Iadd.and for processing substantially only the portion of the signals representing the particular fluorescence to measure the particular fluorescence.Iaddend..

6. The fluorometer of claim 5 additionally including means for producing a scanning of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for forming signals representative of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

7. The fluorometer of claim 6 wherein the means for detecting includes a photosensitive array for detecting the fluorescence emanating from a plurality of spots on the specimen and the means for scanning includes an array control for scanning the photosensitive array to reproduce the detected fluorescence of the different spots on the specimen.

8. The fluorometer of claim 6 wherein the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the fluorescence emanating from the specimen at the different positions.

9. The fluorometer of claim 5 additionally including means responsive .Iadd.substantially only .Iaddend.to the .Iadd.signals representing the particular .Iaddend.fluorescence from the specimen for producing an image of .[.the.]. .Iadd.such .Iaddend.fluorescence .[.and with the controlling means controlling the detecting means within a particular time period by detector responsive means.]. and .Iadd.the detecting means .Iaddend.additionally including means for forming signals including .[.a.]. means for producing a scanning of the fluorescence from the specimen.

10. The fluorometer of claim 9 wherein the means for detecting includes a photosensitive array for detecting the fluorescence emanating from a plurality of spots on the specimen and the means for scanning includes an array control for scanning the photosensitive array to reproduce the detected fluorescence of the different spots on the specimen.

11. The fluorometer of claim 9 wherein the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the fluorescence emanating from the specimen at the different positions.

12. The fluorometer of claim 5 additionally including means responsive to the fluorescence from the specimen for producing an image of the fluorescence and with the controlling means controlling the passage of the image of the fluorescence to the detecting means and additionally including means for forming signals including means for observing or recording of the fluorescence from the specimen.

13. The fluorometer of claim 12 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen.

14. The fluorometer of claim 12 wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

15. The fluorometer of claim 12 wherein the means for detecting includes .[.the.]. .Iadd.a .Iaddend.camera to analyze or record the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

16. The fluorometer of claim 5 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.and .[.with the controlling means controlling the detecting means within a particular time period by detector responsive means and.]. additionally including means for forming signals including means for observing or recording .Iadd.substantially only .Iaddend.the .Iadd.particular .Iaddend.fluorescence from the specimen.

17. The fluorometer of claim 16 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen.

18. The fluorometer of claim 16 wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

19. The fluorometer of claim 5 wherein the means for producing the burst of concentrated light energy is a laser.

20. The fluorometer of claim 5 wherein the means for producing the burst of concentrated light energy is a continuous wave source directing light energy through a modulator.

21. The fluorometer of claim 20 wherein the cubic crystal of the class T_d is cuprous chloride.

22. The fluorometer of claim 20 wherein the electro-optic modulator is formed from at least one of the following group of materials:

CuCl (Cuprous chloride)

CuBr (Cuprous bromide)

CuI (Cuprous iodide)

ZnS (Zinc sulfide)

ZnSe (Zinc selenide)

ZnTe (Zinc telluride)

(CH₂)₆ N₄ (Hexamine or Hexamethylenetetramine)

(Na, Ca)_8-4 (SO₄)_2-1 [(AlSiO₄)₆ ] (Hauynite)

GaP (Gallium phosphide)

Bi₄ (GeO₄)₃ (Bismuth germanate)

NaClO₃ (Sodium chlorate)

BaTiO₃ (Barium titanate)

SrTiO₃ (Strontium titanate)

KTaO₃ (Potassium tantalate)

KTa_x Nb_1-x O₃ (Potassium tantalate niobate)

23. The fluorometer of claim 5 wherein the time period .[.is relatively long and.]. has an ending time sufficiently long to have substantially all of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.decayed .Iadd.at the end of such time period .Iaddend.to a low value relative to the original value of .[.fluorescence.]. .Iadd.such fluorescences.Iaddend..

24. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurements of the particular fluorescence from the measurements of the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the concentrated light energy for directing the concentrated light energy toward the specimen to produce a fluorescence from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.for detecting .[.the fluorescence.]. .Iadd.such fluorescences .Iaddend.and for producing signals in accordance with .[.the fluorescence.]. .Iadd.such detection.Iaddend.,

means coupled to the detecting means for controlling the detecting means within a particular time period .Iadd.after the burst of the concentrated light energy .Iaddend.to optimize the detection of the particular fluorescence and wherein the controlling means is an electro-optic modulator formed by a cubic crystal of the class T_d, the particular fluorescence and the background fluorescence occurring during the particular time period,

.Iadd.the particular fluorescence and the background fluorescence having individual decays and the decay of the particular fluorescence being different from the decay of the background fluorescence, .Iaddend.

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the above means to sequence the detection of the fluorescence within the particular time period after the production of the burst of concentrated light energy, and

means coupled to the .[.detection.]. .Iadd.detecting .Iaddend.means for .[.forming signals representative of the particular fluorescence from the specimen.]. .Iadd.distinguishing the decay of the particular fluorescence during the particular time period from the decay of the background fluorescence during the particular time period and for thereby determining the characteristics substantially only of the particular fluorescence in the specimen.Iaddend..

25. The fluorometer of claim 24 wherein the cubic crystal of the class T_d is cuprous chloride.

26. The fluorometer of claim 24 additionally including means responsive to the fluorescence from the specimen for producing an image of the fluorescence and with the controlling means controlling the passage of the image of the fluorescence to the detecting means and with the means for forming signals including a means for producing a scanning of the fluorescence from the specimen.

27. The fluorometer of claim 26 wherein the means for detecting includes a photosensitive array for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen and the means for scanning includes an array control for scanning the photosensitive array to reproduce the detected .[.fluorescence.]. .Iadd.fluorescences .Iaddend.of the different spots on the specimen.

28. The fluorometer of claim 26 wherein the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen at the different positions.

29. The fluorometer of claim 24 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of the .[.fluorescence and the controlling means including detector responsive means for controlling the detecting means within a particular time period.]. .Iadd.fluorescences .Iaddend.and the means for .[.forming.]. .Iadd.producing .Iaddend.signals including means for producing a scanning of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

30. The fluorometer of claim 29 wherein the means for detecting includes a photosensitive array for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen and the means for scanning includes an array control for scanning the photosensitive array to reproduce the detected .[.fluorescence.]. .Iadd.fluorescences .Iaddend.of the different spots on the specimen.

31. The fluorometer of claim 29 wherein the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen at the different positions.

32. The fluorometer of claim 24 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.and with the controlling means controlling the passage of the image of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.to the detecting means and with the means for forming signals including means for observing or recording .[.of.]. the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

33. The fluorometer of claim 32 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen.

34. The fluorometer of claim 32 wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

35. The fluorometer of claim 32 wherein the means for detecting includes a camera to analyze or record the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

36. The fluorometer of claim 24 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of the .[.fluorescence and with the controlling means controlling the detecting means within a particular time period by detector responsive means.]. .Iadd.fluorescences .Iaddend.and with the means for forming signals including means for observing or recording of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

37. The fluorometer of claim 36 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen.

38. The fluorometer of claim 36 wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

39. The fluorometer of claim 24 wherein the means for producing the burst of concentrated light energy is a laser.

40. The fluorometer of claim 24 wherein the means for producing the burst of concentrated light energy is a continuous wave source directing light energy through a modulator.

41. The fluorometer of claim 24 wherein the .Iadd.particular .Iaddend.time period has a beginning time and an ending time defining a sufficiently short time to enhance the detection of the particular fluorescence relative to the .[.total.]. .Iadd.background .Iaddend.fluorescence.

42. The fluorometer of claim 24 additionally including data processing means responsive to the signals produced by the detecting means for analysing the signals to enhance the portion of the signals representing the particular fluorescence relative to the portion of the signals representing the .[.remaining.]. .Iadd.background .Iaddend.fluorescence.

43. The fluorometer of claim 42 wherein the time period .[.is relatively long and.]. has an ending time sufficiently long to have substantially all of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.decayed to a low value relative to the original value of .[.fluorescence.]. .Iadd.fluorescences.Iaddend..

44. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurements of the particular fluorescence from the measurements of the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the concentrated light energy for directing the concentrated light energy toward the specimen to produce .[.a fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.and for .[.producing.]. .Iadd.analyzing the detected fluorescences as a function of time to produce .Iaddend.signals .[.in accordance with the fluorescence.]. .Iadd.distinguishing the particular fluorescence from the background fluorescence and representing substantially only the particular fluorescence.Iaddend.,

means coupled to the detecting means for controlling the detecting means within a particular time period.Iadd., after the burst of the concentrated light energy, .Iaddend.to optimize the .[.detection of.]. .Iadd.analysis to distinguish .Iaddend.the particular fluorescence .Iadd.from the background fluorescence .Iaddend.and wherein the controlling means is an electro-optic modulator formed from at least one of the following group of materials:

CuCl (Cuprous chloride)

CuBr (Cuprous bromide)

CuI (Cuprous iodine)

ZnS (Zinc sulfide)

ZnSe (Zinc selenide)

ZnTe (Zinc telluride)

(CH₂)₆ N₄ (Hexamine or hexamethylenetetramine)

(Na, Ca)_8-4 (SO₄)_2-1 [(AlSiO₄)₆ ](Hauynite)

GaP (Gallium phosphide)

Bi₄ (GeO₄)₃ (Bismuth germanate)

NaClO₃ (Sodium chlorate)

BaTiO₃ (Barium titanate)

SrTiO₃ (Strontium titanate)

KTaO₃ (Potassium tantalate)

KTa_x Nb_1-x O₃ (Potassium tantalate niobate)

45. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurements of the particular fluorescence from the measurements of the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the concentrated light energy for directing the concentrated light energy toward the specimen to produce .[.a fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence from the specimen .Iaddend.for detecting .[.the fluorescence.]. .Iadd.such fluorescences .Iaddend.and for producing signals in accordance with .[.the fluorescence.]. .Iadd.such detection.Iaddend.,

means coupled to the detecting means for controlling the detecting means within a particular time period.Iadd., after the burst of the concentrated light energy, .Iaddend.to optimize the detection of the particular fluorescence .Iadd.relative to the background fluorescence.Iaddend.,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the above means to sequence the detection of the .Iadd.produced .Iaddend.fluorescence within the particular time period after the production of the burst of concentrated light energy, and

means coupled to the .[.detection.]. .Iadd.detecting .Iaddend.means for forming signals representative of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen, and

data processing means responsive to the signals produced by the .[.detecting.]. .Iadd.signal-forming .Iaddend.means .Iadd.during the particular time period .Iaddend.for .[.analysing the signals to enhance.]. .Iadd.enhancing .Iaddend.the portion of the signals representing the particular fluorescence relative to the portion of the signals representing the .[.remaining.]. .Iadd.background .Iaddend.fluorescence, and wherein the data processing means includes first means for differentiating the signals produced by the detecting means at a number of time points .Iadd.during the particular time period .Iaddend.to produce a plurality of individual time point signals, second means for integrating the signals produced by the detecting means over a .[.corresponding.]. number of time intervals .Iadd.during the particular time period .Iaddend.to produce a plurality of individual time integrated signals and third means responsive to the signals from the .[.detecting.]. .Iadd.signal-forming .Iaddend.means, the individual time point signals and the individual time integrated signals .Iadd.during the particular time period .Iaddend.for operating upon these signals in a particular relationship to .Iadd.distinguish the particular fluorescence from the background fluorescence and to .Iaddend.determine .Iadd.substantially only .Iaddend.the particular fluorescence from the specimen during the particular .Iadd.time .Iaddend.period as a result of the burst of concentrated light energy.

46. The fluorometer of claim 45 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.and with the controlling means controlling the passage of the image of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.to the detecting means and with the means for forming signals including a means for producing a scanning of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

47. The fluorometer of claim 46 wherein the means for detecting includes a photosensitive array for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen and the means for scanning includes an array control for scanning the photosensitive array to reproduce the detected .[.fluorescence.]. .Iadd.fluorescences .Iaddend.of the different spots on the specimen.

48. The fluorometer of claim 46 wherein the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen at the different position.

49. The fluorometer of claim 45 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of .[.the fluorescence and the controlling means including detector responsive means for controlling the detecting means within a particular time period.]. .Iadd.such fluorescences .Iaddend.and the means for forming signals including means for producing a scanning of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

50. The fluorometer of claim 49 wherein the means for detecting includes a photosensitive array for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen and the means for scanning includes an array control for scanning the photosensitive array to reproduce the detected .[.fluorescence.]. .Iadd.fluorescences .Iaddend.of the different spots on the specimen.

51. The fluorometer of claim 49 wherein the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen at the different positions.

52. The fluorometer of claim 45 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.and with the controlling means controlling the passage of the image of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.to the detecting means and with the means for forming signals including means for observing or recording of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

53. The fluorometer of claim 52 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen.

54. The fluorometer of claim 52 wherein the means for detecting includes an image tube to reproduce the pattern of fluorescence emanating from a plurality of spots on the specimen.

55. The fluorometer of claim 52 wherein the means for detecting includes a camera to analyze or record the fluorescence emanating from a plurality of spots on the specimen.

56. The fluorometer of claim 45 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of the .[.fluorescence and with the controlling means controlling the detecting means within a particular time period by detector responsive means.]. .Iadd.fluorescences .Iaddend.and with the means for forming signals including means for observing or recording .[.of.]. the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

57. The fluorometer of claim 56 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen.

58. The fluorometer to claim 56 wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

59. The fluorometer of claim 45 wherein the means for producing the burst of concentrated light energy is a laser.

60. The fluorometer of claim 45 wherein the means for producing the burst of concentrated light energy is a continuous wave source directing light energy through a modulator.

61. The fluorometer of claim 45 wherein the means for controlling the detecting means within a particular time period is an electro-optic modulator.

62. The fluorometer of claim 45 wherein the .Iadd.particular .Iaddend.time period has a beginning time and an ending time defining a sufficiently short time to enhance the detection of the particular fluorescence relative to the .[.total .]. .Iadd.detection of the background .Iaddend.fluorescence.

63. The fluorometer of claim 45 wherein the .Iadd.particular .Iaddend.time period .[.is relatively long and.]. has an ending time sufficiently long to have substantially all of the fluorescence decayed to a low value relative to the original value of fluorescence.

64. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurement of the particular fluorescence from the measurement of the background fluorescence.Iaddend., including,

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the burst of concentrated light energy for directing the burst of concentrated light energy toward the specimen to produce a fluorescence from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen for detecting .[.the fluorescence.]. .Iadd.such fluorescences .Iaddend.and for producing signals in accordance with such detection,

means coupled to the detecting means for obtaining a controlled operation of the detecting means during a particular time period .Iadd.after the burst of the concentrated light energy .Iaddend.to optimize the detection of the particular fluorescence and wherein the .Iadd.particular .Iaddend.time period has a beginning time and an ending time to enhance the detection of the .Iadd.portion of the .Iaddend.signals representing the particular fluorescence relative to the portion of the signals representing the .[.remaining.]. .Iadd.background .Iaddend.fluorescence, the beginning time of the particular time period being a first particular time after the burst of .Iadd.the .Iaddend.concentrated light energy and after the production of the particular fluorescence .Iadd.and the background fluorescence.Iaddend., and the ending time being a second particular time after the first particular time and during the production of the particular fluorescence from the specimen as a result of the burst of concentrated light energy,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the detecting means to sequence the detection of the .Iadd.particular .Iaddend.fluorescence during the particular time period as a result of the production of the burst of concentrated light energy, and

data processing means responsive to the signals produced by the detecting means during the particular time period for .[.analysing the signals to enhance.]. .Iadd.distinguishing .Iaddend.the portion of the signals representing the particular fluorescence during the particular time .[.interval relative to.]. .Iadd.period from .Iaddend.the portion of the signals representing the .[.remaining.]. .Iadd.background .Iaddend.fluorescence during the particular time .[.interval.]. .Iadd.period and for producing signals representing substantially only the particular fluorescence.Iaddend..

65. A fluorometer as set forth in claim 64 wherein the detecting means includes means for scanning the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen to obtain the production of the signals representative of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen as a result of the production of the burst of concentrated light energy.

66. The fluorometer of claim 65 wherein the means for detecting includes a photosensitive array for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen and the means for scanning includes an array control for scanning the photosensitive array to reproduce the detected .[.fluorescence.]. .Iadd.fluorescences .Iaddend.of the different spots on the specimen.

67. The fluorometer of claim 65 wherein the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen at the different positions.

68. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurements of the particular fluorescence from the measurements of the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the concentrated light energy for directing the concentrated light energy toward the specimen to produce .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence from the specimen .Iaddend.for detecting .[.the fluorescence.]. .Iadd.such fluorescences .Iaddend.and for producing signals in accordance with .[.the fluorescence.]. .Iadd.such detection.Iaddend.,

means coupled to the detecting means for controlling the detecting means within a particular time period.Iadd., after the burst of the concentrated light energy, .Iaddend.to optimize the detection of the particular fluorescence,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the above means to sequence the detection of the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence from the specimen .Iaddend.within the particular time period after the production of the burst of .Iadd.the .Iaddend.concentrated light energy, .Iadd.and .Iaddend.

data processing means responsive to the signals produced by the detecting means .Iadd.during the particular time period .Iaddend.for .[.analysing the signals to enhance.]. .Iadd.enhancing .Iaddend.the portion of the signals representing the particular fluorescence .Iadd.during the particular time period .Iaddend.relative to the portion of the signals representing the .[.remaining.]. .Iadd.background .Iaddend.fluorescence .Iadd.during the particular time period.Iaddend., and

the data processing means including first means for differentiating the signals produced by the detecting means at a number of time points .Iadd.during the particular time period .Iaddend.to produce a plurality of individual time point signals, second means for integrating the signals produced by the detecting means over a .[.corresponding.]. number of time intervals .Iadd.during the particular time period .Iaddend.to produce a plurality of individual time integrated signals and third means responsive to the signals from the detecting means, the individual time point signals and the individual time integrated signals .Iadd.during the particular time period .Iaddend.for producing signals representing .Iadd.substantially only .Iaddend.the particular fluorescence.

69. The fluorometer of claim 68 additionally including means for producing a scanning of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for forming signals representative of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

70. The fluorometer of claim 69 wherein the means for detecting includes a photosensitive array for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen and the means for scanning includes an array control for scanning the photosensitive array to reproduce the detected .[.fluorescence.]. .Iadd.fluorescences .Iaddend.of the different spots on the specimen.

71. The fluorometer of claim 69 wherein the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen at the different positions.

72. The fluorometer of claim 69 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of the .[.fluorescence and the controlling means including detector responsive means for controlling the detecting means within a particular time period.]. .Iadd.fluorescences.Iaddend..

73. The fluorometer of claim 72 wherein the means for detecting includes a photosensitive array for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen and the means for scanning includes an array control for scanning the photosensitive array to reproduce the detected .[.fluorescence.]. .Iadd.fluorescences .Iaddend.of the different spots on the specimen.

74. The fluorometer of claim 72 wherein the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen at the different positions.

75. The fluorometer of claim 68 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.and with the controlling means controlling the passage of the image of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.to the detecting means and additionally including means for forming signals including means for observing or recording .[.of.]. the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

76. The fluorometer of claim 75 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

77. The fluorometer of claim 75 wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

78. The fluorometer of claim 75 wherein the means for detecting includes .[.the human eye or.]. a camera to analyze or record the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

79. The fluorometer of claim 68 additionally including means responsive to the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen for producing an image of the .[.fluorescence and with the controlling means controlling the detecting means with a particular time period by detector responsive means.]. .Iadd.fluorescences .Iaddend.and additionally including means for forming signals including means for observing or recording .[.of.]. the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen.

80. The fluorometer of claim 79 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen.

81. The fluorometer of claim 79 wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

82. The fluorometer of claim 68 wherein the means for producing the burst of concentrated light energy is a laser.

83. The fluorometer of claim 68 wherein the means for producing the burst of concentrated light energy is a continuous wave source directing light energy through a modulator.

84. The fluorometer of claim 68 wherein the means for controlling the passage of the image is an electro-optic modulator.

85. The fluorometer of claim 84 wherein the electro-optic modulator is formed by a cubic crystal of the class T_d.

86. The fluorometer of claim 85 wherein the cubic crystal of the class T_d is cuprous chloride.

87. The fluorometer of claim 85 wherein the electro-optic modulator is formed from at least one of the following group of materials:

CuCl (Cuprous chloride)

CuBr (Cuprous bromide)

CuI (Cuprous iodide)

ZnS (Zinc sulfide)

ZnSe (Zinc selenide)

ZnTe (Zinc telluride)

(CH₂)₆ N₄ (Hexamine or Hexamethylenetetramine)

(Na, Ca)_8-4 (SO₄)_2-1 [(AlSiO₄)₆ ] (Hauynite)

GaP (Gallium phosphide)

Bi₄ (GeO₄)₃ (Bismuth germanate)

NaClO₃ (Sodium chlorate)

BaTiO₃ (Barium titanate)

SrTiO₃ (Strontium titanate)

KTaO₃ (Potassium tantalate) KTa_x Nb_1-x O₃ (Potassium tantalate niobate)

88. The fluorometer of claim 68 wherein the .Iadd.particular .Iaddend.time period .[.is relatively long and.]. has an ending time sufficiently long to have substantially all of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.decayed to a low value relative to the original value of .[.fluorescence.]. .Iadd.fluorescences.Iaddend..

89. The fluorometer of claim 68 wherein the .Iadd.particular .Iaddend.time period has a beginning time and an ending time defining a sufficiently short time to enhance the detection of the signals representing the particular fluorescence relative to the portion of the signals representing the .[.remaining.]. .Iadd.background .Iaddend.fluorescence.

90. The fluorometer of claim 68 wherein the .Iadd.particular fluorescence has a characteristic decay time and .Iaddend.first means produces the plurality of individual time point signals .[.produces the plurality of individual time point signals.]. for a specimen not carrying a fluorescent label, said specimen being not necessarily of the same size and disposition as that of the specimen carrying the fluorescent label but consisting of the same material but without .Iadd.the .Iaddend.added fluorescent label, and the third means extracts from the individual time point signals and the derivatives at the corresponding time points and the characteristics decay time of the particular .[.fluorescence.]. .Iadd.fluorescences .Iaddend.the emission intensity representing the particular fluorescence only and being separated from the background emission.

92. The fluorometer of claim 71 wherein the means for detecting includes a photosensitive array for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen and the means for producing signals includes an array control for scanning the photosensitive array to obtain a reproduction of the detected .[.fluorescence.]. .Iadd.fluorescences .Iaddend.of the different spots on the specimen.

91. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing between the measurement of the particular fluorescence and the measurement of the background fluorescence.Iaddend., including

.Iadd.the particular fluorescence having a different decay time than the background fluorescence, .Iaddend.

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the burst of concentrated light energy for directing the burst of concentrated light energy toward the specimen to produce a fluorescence from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen for detecting .[.the fluorescence.]. .Iadd.such fluorescences .Iaddend.and for producing signals in accordance with such detection,

means coupled to the detecting means for obtaining a controlled operation of the detecting means in a particular time period to optimize the detection of the particular fluorescence wherein the .Iadd.particular .Iaddend.time period has a beginning time and an ending time to enhance the detection of the particular fluorescence relative to the .[.total.]. .Iadd.background .Iaddend.fluorescence, the beginning time occurring a first particular time after the burst of concentrated light energy and during the production of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.and the ending time occurring a second particular time after the first particular time and during the production of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen as a result of the burst of concentrated light energy,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the detecting means to sequence the detection of the fluorescence during the particular time period as a result of the production of the burst of concentrated light energy, .Iadd.and .Iaddend.

means coupled to the detecting means for .Iadd.distinguishing the particular decay time of the particular fluorescence during the particular time period from the decay time of the background fluorescence during the particular time period and for .Iaddend.forming signals representative .Iadd.substantially only .Iaddend.of the particular fluorescence from the specimen, and

means responsive to the .[.detection of.]. .Iadd.signals representing substantially only .Iaddend.the particular fluorescence from the specimen during the particular time period for producing an image of .Iadd.substantially only .Iaddend.the particular fluorescence during the particular time interval.

93. The fluorometer of claim 91 including, a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen at the different positions.

94. A fluorometer for measuring the .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurement of the particular fluorescence from the measurement of the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the burst of concentrated light energy for directing the burst of concentrated light energy toward the specimen to produce a fluorescence from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen for detecting .[.the fluorescence.]. .Iadd.such fluorescences .Iaddend.and for producing signals in accordance with such detection,

means coupled to the detecting means for obtaining a controlled operation of the detecting means during a particular time period.Iadd., after the burst of the concentrated light energy, .Iaddend.to optimize the detection of the particular fluorescence wherein the particular time period has a beginning time and an ending time to enhance the detection of the particular fluorescence relative to the .[.total.]. .Iadd.background .Iaddend.fluorescence, the beginning time and the ending time occurring during the production of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen .[.as a result of.]. .Iadd.after .Iaddend.the burst of concentrated light energy,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the detecting means to sequence the detection of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.during the particular time period as a result of the production of the burst of concentrated light energy,

means coupled to the detecting means for .Iadd.distinguishing the signals representing the particular fluorescence during the particular time period from the signals representing the background fluorescence during the particular time period and for .Iaddend.forming signals representative .Iadd.substantially only .Iaddend.of the particular fluorescence from the specimen during the particular time period,

means responsive to the .Iadd.signals representing substantially only the particular .Iaddend.fluorescence from the specimen for producing an image of .Iadd.substantially only .Iaddend.the particular fluorescence during the particular time period.

95. The fluorometer of claim 94 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen.

96. The fluorometer of claim 94 wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

97. The fluorometer of claim 94 wherein the means for detecting includes the camera to analyze or record the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

98. The fluorometer of claim 94 wherein the means for producing the burst of concentrated light energy is a continuous wave source directing light energy through a modulator.

99. The fluorometer of claim 94 wherein the means for controlling the detecting means within a particular time period is an electro-optic modulator.

100. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurement of the particular fluorescence from the measurement of the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the burst of concentrated light energy for directing the burst of concentrated light energy toward the specimen to produce .[.a fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen for detecting .[.the fluorescence.]. .Iadd.such fluorescences .Iaddend.and for producing signals in accordance with such detection,

means coupled to the detecting means for obtaining a controlled operation of the detecting means during a particular time period.Iadd., after the burst of the concentrated light energy, .Iaddend.to optimize the detection of the particular fluorescence during the particular time period, the particular time period having a beginning time and an ending time to enhance the detection of the particular fluorescence relative to the .Iadd.background .Iaddend.fluorescence .[.including the particular flourescence.]., the beginning time and ending time occurring during the production of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen as a result of the production of the burst of concentrated energy,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the detecting means to sequence the detection of the fluorescence during the particular time period as a result of the production of the burst of concentrated light energy,

.Iadd.data processing .Iaddend.means coupled to the detecting means for .[.forming.]. .Iadd.differentiating the detected fluorescence, and integrating the detected fluorescence, at progressive instants of time and for processing the results of such differentiation and integration to form .Iaddend.signals representative .Iadd.substantially only .Iaddend.of the particular fluorescence from the specimen, and

means responsive to the signals from the .[.detecting.]. .Iadd.data processing .Iaddend.means during the particular time period .Iadd.in representation of substantially only the particular fluorescence from the specimen .Iaddend.for producing an image of .Iadd.substantially only .Iaddend.the particular fluorescence during the particular time period.

101. The fluorometer of claim 100 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen.

102. The fluorometer of claim 100 wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

103. The fluorometer of claim 100 wherein the means for producing the burst of concentrated light energy is a laser.

104. The fluorometer of claim 100 additionally including data processing means responsive to the signals produced by the detecting means for analysing the signals to enhance the portion of the signals representing the particular fluorescence relative to the portion of the signals representing the .[.remaining.]. .Iadd.background .Iaddend.fluorescence.

105. A fluorometer as set forth in claim 100, including,

means for recording the particular fluorescence from the specimen during the particular time period as a result of the burst of concentrated light energy.

106. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in a specimen .Iadd.and for distinguishing the measurement of the particular fluorescence from the measurement of the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy of excite substantially all of the particular fluorophore molecules,

means responsive to the burst of concentrated light energy for directing the burst of concentrated light energy toward the specimen to produce a fluorescence from the specimen including the particular fluorescence,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen for detecting .[.the fluorescence.]. .Iadd.such fluorescences .Iaddend.and for producing signals in accordance with such detection,

means coupled to the detecting means for obtaining a controlled operation of the detecting means during a particular time period .Iadd.after the burst of concentrated light energy .Iaddend.to optimize the detection of the particular fluorescence during the particular time period.Iadd., .Iaddend.the particular time period having a beginning time and an ending time to enhance the detection of the .Iadd.portion of the .Iaddend.signals representing the particular fluorescence during the particular time period relative to the portion of the signals representing the .[.remaining.]. .Iadd.background .Iaddend.fluorescence during the particular time period, the beginning time and the ending time occurring during the production of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen as a result of the burst of concentrated light energy,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the detecting means to sequence the detection of the fluorescence from the specimen during the particular time period as a result of the production of the burst of concentrated light energy,

data processing means responsive to the signals produced by the detecting means for analyzing the signals to enhance the portion of the signals representing the particular fluorescence during the particular time interval relative to the portion of the signals representing the fluorescence during the particular time interval other than the particular fluorescence, and

means responsive to the .[.signal.]. .Iadd.signals .Iaddend.from the data processing means for producing an image of the particular fluorescence.

107. The fluorometer of claim 106 wherein the means for detecting includes a photosensitive array for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen and means are included for scanning the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen and the means for scanning includes an array control for scanning the photosensitive array to reproduce the detected .[.fluorescence.]. .Iadd.fluorescences .Iaddend.of the different spots on the specimen.

108. The fluorometer of claim 106 wherein means are included for scanning the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen and the means for scanning includes a stepping stage for supporting the specimen and for moving the specimen to a plurality of different positions for providing a detection of the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen at the different positions.

109. A fluorometer as set forth in claim 106 wherein the

means for forming signals includes means for scanning the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.produced from the specimen during the particular time period as a result of the burst of concentrated light energy.

112. The fluorometer of claim .[.106.]. .Iadd.110 .Iaddend.wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

110. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurement of the particular fluorescence from the measurement of the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the burst of concentrated light energy for directing the burst of concentrated light energy toward the specimen to produce .[.fluorescence from the specimen including.]. the particular fluorescence .Iadd.and the background fluorescence from the specimen.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen for detecting such .[.fluorescence.]. .Iadd.fluorescences .Iaddend.and for producing signals in accordance with such detection,

means coupled to the detecting means for obtaining a controlled operation of the detecting means within a particular time period.Iadd., after the burst of the concentrated light energy,.Iaddend.to optimize the detection of the particular fluorescence wherein the particular time period has a beginning time and an ending time to enhance the detection of .[.the signals representing.]. the particular fluorescence during the particular time period relative to the .[.portion of the signals representing.]. .Iadd.detection of .Iaddend.the .Iadd.background .Iaddend.fluorescence during the particular time period .[.other than the particular fluorescence.]., the beginning and ending times occurring during the production of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen .[.as a result of.]. .Iadd.after .Iaddend.the burst of concentrated .Iadd.light .Iaddend.energy,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the detecting means to sequence the detection of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen during the particular time period as a result of the production of the burst of concentrated .Iadd.light .Iaddend.energy, .Iadd.and .Iaddend.

data processing means responsive to the signals produced by the detecting means during the particular time period for .[.analyzing the signals to enhance.]. .Iadd.enhancing as a function of time .Iaddend.the portion of the signals representing the particular fluorescence during the particular time period relative to the portion of the signals representing the .Iadd.background .Iaddend.fluorescence during the particular time period .[.other than particular fluorescence.]. .Iadd.and for producing signals representing substantially only the particular fluorescence during the particular time period.Iaddend., and

means responsive to the signals from the data processing means .Iadd.in representation of substantially only the particular fluorescence .Iaddend.for producing an image of the .Iadd.particular .Iaddend.fluorescence.

111. The fluorometer of claim 110 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from the specimen.

113. The fluorometer of claim 110 wherein the means for detecting includes a camera to analyze or record the .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

114. The fluorometer of claim 110 wherein the means for producing the burst of concentrated light energy is a continuous wave source directing light energy through a modulator.

115. A fluorometer as set forth in claim 110, including,

means for recording the particular fluorescence produced from the specimen during the particular time period as a result of the burst of concentrated light energy.

116. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurement of the particular fluorescence from the measurement of the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the burst of concentrated light energy for directing the burst of concentrated light energy toward the specimen to produce .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen for detecting such .[.fluorescence.]. .Iadd.fluorescences .Iaddend.and for producing signals in accordance with such detection,

means coupled to the detecting means for obtaining a controlled operation of the detecting means within a particular time period .Iadd.after the burst of the concentrated light energy .Iaddend.to optimize the detection of the particular fluorescence wherein the .Iadd.particular .Iaddend.time period has a beginning time and an ending time to enhance the detection of .[.the signals representing.]. the particular fluorescence during the particular time interval relative to the .[.portion of the signals representing.]. .Iadd.detection of .Iaddend.the .Iadd.background .Iaddend.fluorescence during the particular time period .[.other than the particular fluorescence.]., the beginning and ending times occurring during the production of the particular fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen .[.as a result of.]. .Iadd.after .Iaddend.the burst of concentrated light energy,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the detecting means to sequence the detection of the fluorescence within the particular time period as a result of the production of the burst of concentrated light energy,

data processing means responsive to the signals produced by the detecting means during the particular time period for .[.analysing the signals to enhance.]. .Iadd.enhancing .Iaddend.the portion of the signals representing the particular fluorescence during the particular time period relative to the portion of the signals representing the .Iadd.background .Iaddend.fluorescence during the particular time period .[.other than the particular fluorescence.]., and

means responsive to the enhanced signals from the data processing means for producing an image of .Iadd.substantially .Iaddend.the particular fluorescence during the particular time period.

117. The fluorometer of claim 116 wherein the means for detecting includes a photodetector for detecting the .[.fluorescence.]. .Iadd.fluorescences emanating from the specimen.

118. The fluorometer of claim 116 wherein the means for detecting includes an image tube to reproduce the pattern of .[.fluorescence.]. .Iadd.fluorescences .Iaddend.emanating from a plurality of spots on the specimen.

119. The fluorometer of claim 116 wherein the means for producing the burst of concentrated light energy is a laser.

120. A fluorometer as set forth in claim 116, including,

means for recording the particular fluorescence produced from the specimen during the particular time period as a result of the burst of concentrated light energy.

121. A fluorometer for measuring .Iadd.the fluorescence from a specimen having a background fluorescence and .Iaddend.a particular fluorescence emanating from particular fluorophore molecules in .[.a.]. .Iadd.the .Iaddend.specimen .Iadd.and for distinguishing the measurement of the particular fluorescence from the measurement of the background fluorescence.Iaddend., including

means for producing a burst of concentrated light energy having a pulse time short compared to the decay time of the particular fluorescence and having sufficient energy to excite substantially all of the particular fluorophore molecules,

means responsive to the burst of concentrated light energy for directing the burst of concentrated light energy toward the specimen to produce .[.fluorescence.]. .Iadd.fluorescences .Iaddend.from the specimen including the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means responsive to the .Iadd.particular .Iaddend.fluorescence .Iadd.and the background fluorescence .Iaddend.from the specimen for detecting such .[.fluorescence.]. .Iadd.fluorescences .Iaddend.and for producing signals in accordance with such detection,

means coupled to the detecting means for obtaining a controlled operation of the detecting means within a particular time period.Iadd., after the burst of concentrated light energy, .Iaddend.to optimize the detection of the particular fluorescence wherein the particular time period has a beginning time and an ending time to enhance the detection of .[.the signals representing.]. the particular fluorescence during the particular time .Iadd.period .Iaddend.relative to the .[.portion of the signals representing.]. .Iadd.detection of .Iaddend.the .Iadd.background .Iaddend.fluorescence during the particular time period .[.other than the particular fluorescence.]..Iadd., .Iaddend.the beginning and ending times occurring during the production of the particular fluorescence .Iadd.and the background fluorescence.Iaddend.,

means coupled to the burst producing means, to the detecting means and to the controlling means for timing the operation of the detecting means to sequence the detection of the fluorescence during the particular time period .[.as a result of.]. .Iadd.after .Iaddend.the production of the burst of concentrated light energy, .Iadd.and .Iaddend.

data processing means responsive to the signals produced by the detecting means during the particular time period for .[.analyzing the signals to enhance.]. .Iadd.enhancing .Iaddend.the portion of the signals representing the particular fluorescence during the particular time period relative to the portion of the signals representing the .Iadd.background .Iaddend.fluorescence during the particular time period .[.other than the particular fluorescence.]., and

the means for controlling the .[.passage of the image.]. .Iadd.detection of the fluorescences .Iaddend.constituting an electro-optic modulator formed by a cubic crystal of the class T_d.

122. The fluorometer of claim 121 wherein the cubic crystal of the class T_d is cuprous chloride.

123. The fluorometer of claim 121 wherein the electro-optic modulator is formed from at least one of the following group of materials:

CuCl (Cuprous chloride)

CuBr (Cuprous bromide)

CuI (Cuprous iodide)

ZnS (Zinc sulfide)

ZnSe (Zinc selenide)

ZnTe (Zinc telluride)

(CH₂)₆ N₄ (Hexamine or Hexamethylenetetramine)

(Na, Ca)_8-4 (SO₄)_2-1 [(AlSiO₄)₆ ]](Hauynite)

GaP (Gallium phosphide)

Bi₄ (GeO₄)₃ (Bismuth germanate)

NaClO₃ (Sodium chlorate)

BaTiO₃ (Barium titanate)

SrTiO₃ (Strontium titanate)

KTaO₃ (Potassium tantalate)

KTa_x Nb_1-x O₃ (Potassium tantalate niobate)