System and method for controlled intensity illumination in a bioanalysis or other system

• US 9,658,160 B2
• Filed: 04/11/2016
• Issued: 05/23/2017
• Est. Priority Date: 01/14/2011
• Status: Active Grant

• Current Assignee: Lumencor, Inc.
• Original Assignee: Lumencor, Inc.
• Inventors: Jaffe, Claudia B, Jaffe, Steven M, Larsen, David
• Primary Examiner(s): HAMMOND, DEDEI KHALIEAH
• Application Number:
  US15/095,397
• Publication Number:
  US 20160223461A1
• Time in Patent Office: 407 Days
• Field of Search: None
• CPC Class Codes: F21V 23/0457 the sensor sensing the operating status of the lighting device, e.g. to detect failure of a light source or to provide feedback to the device
• CPC Class Codes: G01J 1/4257 applied to monitoring the characteristics of a beam, e.g. laser beam, headlamp beam monitoring arrangements for lasers in general H01S3/0014
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence
• CPC Class Codes: G01N 21/6408 with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 2201/061 Sources
• CPC Class Codes: H05B 45/22 using optical feedback

Abstract

A system and method for metered dosage illumination in a bioanalysis or other system. In accordance with an embodiment, an illumination system or subsystem is described that can provide optimized amounts of excitation light within the short exposure times necessary to measure fast biological activity. The amount of light can be precisely measured to provide quantitative results. The light flux can also be precisely controlled to generate only a prescribed minimum amount of light, in order to reduce adverse lighting effects on both fluors and samples. Although the examples herein illustrate the providing of metered dosage illumination in the context of a bioanalysis system, the techniques can be similarly used to provide metered dosage illumination in the context of other types of system. In accordance with various embodiments, the technique is particularly useful in any quality-control, analysis, or assessment-based environment. Typical research and development applications can include quality control, instrument calibration, and light output standardization; while clinical and diagnostics applications can include clinical monitoring, bioassay calibration and control for diagnostics, treatment and or therapeutic evaluation.

Claims

1. A system for providing metered dosage illumination for use in a bioanalysis or other system, comprising:

a light engine including a plurality of color channels wherein each color channel includes an LED light source, a band pass filter, and an electronic circuit to power the LED light source, whereby each color channel provides light of a different specific wavelength band;

an optical system comprising a plurality of reflective elements internal to the system configured to direct light from each color channel into a light beam;

a beam splitter positioned in the light beam and configured to split said light beam into an output light beam for illuminating said sample and a redirected light beam internal to the system;

a light-to-frequency converter positioned internal to the system to receive said redirected light beam from the beam splitter and generate a pulse train having a frequency proportional to an intensity of said redirected light beam;

a counter that maintains a count of pulses in the pulse train when a particular color channel of the plurality of color channels is turned on and turns off the particular color channel when the count of pulses reaches a reference count.

2. The system of claim 1, wherein said plurality of color channels comprises at least four color channels selected from violet, blue, cyan, teal, green, red, and yellow, such that each of the four color channels provides light of a selected intensity and a different specific wavelength band suitable for exciting a fluorescent molecule, and wherein the plurality of color channels includes at least four solid state light sources and at least four band pass filters.

3. The system of claim 1, wherein said plurality of color channels comprises at least five color channels selected from violet, blue, cyan, teal, green, red, and yellow, such that each of the five color channels provides light of a selected intensity and a different specific wavelength band suitable for exciting a fluorescent molecule, and wherein the plurality of color channels includes at least five solid state light sources and at least five band pass filters.

4. The system of claim 1, wherein the system further comprises a camera for imaging a sample exposed to the output light beam and wherein the system gates the camera off when the count of pulses reaches the reference count.

5. The system of claim 1, further comprising:

a camera for measuring fluorescence received from a sample in response to exposure of the sample to the output light beam;

a host computer for triggering an exposure period of said camera; and

wherein the system is configured to end the exposure period of said camera when the count of pulses reaches the reference count.

6. The system of claim 1, wherein the system is used as part of a bioanalysis system for one or more of:

quantitative analyses, temporally fast analyses, fluorescence imaging, gene expression analysis, sequencing, high resolution fluorescence microscopy, fluorescence life time measurements, Fø

rster Resonance Energy Transfer (FRET) Microscopy, and high content screening.

7. The system of claim 1, wherein the system further comprises:

a light detector for quantitative measurement of fluorescent light emitted by a sample in response to exposure of the sample to the output light beam; and

wherein the system is configured to turn on the light detector for quantitative measurements and gate the light detector off when the frequency detector count equals the reference count.

8. The system of claim 1, wherein turning off the particular color channel when the count of pulses reaches a reference count enables the system to control a light flux applied to a sample illuminated by the output beam in order to allow quantitative measurement of fluorescence produced by the sample in response to said light flux.

9. The system of claim 1, wherein turning off the particular color channel when the count of pulses reaches a reference count enables the system to control a light flux applied to a sample illuminated by the output beam in order to reduce photo-bleaching of the sample.

10. The system of claim 1, wherein turning off the particular color channel when the count of pulses reaches a reference count enables the system to control a light flux applied to a live cell sample illuminated by the output beam in order to reduce photo-toxic effects on the live cell sample.

11. A method for providing metered dosage illumination in a bioanalysis or other system, comprising:

providing a light engine comprising,

a plurality of color channels wherein each color channel includes an LED light source, a band pass filter, and an electronic circuit to power the LED light source, whereby each color channel provides light of a different specific wavelength band,

an optical system comprising a plurality of reflective elements internal to the system configured to direct light from each color channel into a light beam,

a beam splitter positioned in the light beam and configured to split said light beam into an output light beam for illuminating said sample and a redirected light beam internal to the system,

a light-to-frequency converter positioned internal to the system to receive said redirected light beam from the beam splitter, and

a counter

turning on a particular color channel of the plurality of color channels;

generating with the light-to-frequency converter a pulse train having a frequency proportional to an intensity of said redirected light beam;

maintaining with the counter a count of pulses in the pulse train; and

turning off the particular color channel when the count of pulses reaches a reference count.

12. The method of claim 11, wherein said plurality of color channels comprises at least four color channels selected from violet, blue, cyan, teal, green, red, and yellow, such that each of the four color channels provides light of a selected intensity and a different specific wavelength band suitable for exciting a fluorescent molecule, and wherein the plurality of color channels includes at least four solid state light sources and at least four band pass filters.

13. The method of claim 11, wherein said plurality of color channels comprises at least five color channels selected from violet, blue, cyan, teal, green, red, and yellow, such that each of the five color channels provides light of a selected intensity and a different specific wavelength band suitable for exciting a fluorescent molecule, and wherein the plurality of color channels includes at least five solid state light sources and at least five band pass filters.

14. The method of claim 11, further comprising:

measuring fluorescence received from a sample in response to exposure of the sample to the output light beam using a camera.

15. The method of claim 11, further comprising:

imaging a sample exposed to the output light beam with a camera; and

gating the camera off when the count of pulses reaches the reference count.

16. The method of claim 11 further comprising:

triggering an exposure period of a camera;

measuring with said camera fluorescence received from a sample in response to exposure of the sample to the output light beam; and

ending the exposure period of said camera when the count of pulses reaches the reference count.

17. The method of claim 11, further comprising:

turning on a light detector for quantitative measurements of fluorescent light emitted by a sample in response to exposure of the sample to the output light beam; and

gating the light detector off when the frequency detector count equals the reference count.

18. The method of claim 11, further comprising controlling a light flux applied to a sample illuminated by the output beam in order to allow quantitative measurement of fluorescence produced by the sample in response to said light flux.

19. The method of claim 11, further comprising controlling a light flux applied to a sample illuminated by the output beam in order to reduce photo-bleaching of the sample.

20. The method of claim 11, further comprising controlling a light flux applied to a live cell sample illuminated by the output beam in order to reduce photo-toxic effects on the live cell sample.