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

• US 9,335,266 B2
• Filed: 05/01/2013
• Issued: 05/10/2016
• 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): Owens, Douglas W
• Assistant Examiner(s): Hammond, Dedei K
• Application Number:
  US13/874,645
• Publication Number:
  US 20130234047A1
• Time in Patent Office: 1,105 Days
• Field of Search: None
• US Class Current: 1/1
• 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 controlled illumination in a bioanalysis or other system where excitation of fluorescent molecules is desirable. In an embodiment, an illumination system is described which can provide excitation light at a controlled intensity to provide quantitative results. In an embodiment, a solid state light engine is described which includes a plurality of color channels each providing light output suitable for exciting a fluorescent molecule, a light to frequency converter which receives a portion of the light output, a counter which maintains a count of a signal from the light to frequency converter, and a light intensity circuit, responsive to the counter, which adjusts the color channels to control the intensity of the light output.

Claims

1. A system for providing light to illuminate a sample comprising:

a light engine including a plurality of color channels wherein each color channel includes,

a solid state light source,

a band pass filter, and

an electronic circuit to power the solid state light source,

whereby each color channel provides light of a different specific wavelength band suitable for exciting a fluorescent molecule;

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

a beam splitter which splits 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 internal to the system which receives said redirected light beam from the beam splitter and generates 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 such that the count of pulses provides a measure of the intensity of light in said redirected light beam; and

a light intensity circuit which monitors the ongoing count of pulses maintained by the counter and adjusts one or more of the plurality of color channels in real time to thereby control intensity of said output light beam for illuminating the sample.

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, further comprising:

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

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; and

a host computer for triggering an exposure period of said camera.

6. The system of claim 1, wherein the light intensity circuit monitors the ongoing count of pulses maintained by the counter and adjusts one or more of the plurality of color channels in real time to thereby maintain constant output light beam intensity.

7. The system of claim 1, wherein the system is used as part of a bioanalysis system.

8. The system of claim 1, wherein the system is used as part of a bioanalysis system used for detection of fluorescent molecules in a biological sample.

9. 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, Fö

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

10. A method comprising:

providing a light engine including a plurality of color channels, wherein each color channel includes, a solid state light source, a band pass filter, and an electronic circuit to power the solid state light source, and whereby each color channel provides light of a different specific wavelength band suitable for exciting a fluorescent molecule;

causing one or more of the color channels to provide light of a different specific wavelength band suitable for exciting a fluorescent molecule;

combining the light from each of the plurality of color channel into a light beam using a plurality of reflective elements;

splitting the light beam using a beamsplitter into a redirected light beam directed at a light to frequency converter internal to the light engine and an output light beam for illuminating a sample;

receiving the redirected light beam from the beam splitter with a light to frequency converter;

generating a pulse train with said light to frequency converter wherein the pulse train has a frequency proportional to an intensity of light in said redirected light beam;

maintaining a count of pulses in the pulse train such that the count of pulses provides a measure of the intensity of light in said redirected of the light beam; and

adjusting one or more of the plurality of color channels in real time in response to the count of pulses to thereby control intensity of said output light beam for illuminating the sample.

11. The method of claim 10, wherein the plurality of color channels includes 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 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.

12. The method of claim 10, wherein the plurality of color channels includes 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 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.

13. The method of claim 10, further comprising:

directing the output light beam at a sample; and

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

14. The method of claim 10, further comprising:

directing the output light beam at a sample;

triggering an exposure period of a camera;

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

15. The method of claim 10, wherein said adjusting step comprises adjusting one or more of the plurality of color channels in real time in response to the count of pulses to thereby maintain constant output light beam intensity.

16. The method of claim 10, further comprising:

directing the output light beam at a sample; and

measuring fluorescence received from the sample in response to exposure of the sample to the output light beam to perform analysis of said sample.

17. The method of claim 10, further comprising:

directing the output light beam at a biological sample; and

measuring fluorescence received from the biological sample in response to exposure of the biological sample to the output light beam to detect fluorescent molecules in the biological sample.

18. The method of claim 10, further comprising:

directing the output light beam at a sample; and

measuring fluorescence received from the sample in response to exposure of the sample to the output light beam to perform analysis of said sample wherein said analysis includes one or more of;

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

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

19. A system for the detection of fluorescent molecules in a biological sample, the system comprising:

a light engine including a plurality of color channels wherein each color channel includes,

a solid state light source,

a band pass filter, and

an electronic circuit to power the solid state light source,

whereby each color channel provides light of a different specific wavelength band suitable for exciting a fluorescent molecule;

an optical system comprising a plurality of reflective elements which direct light from each color channel into a light beam;

a beam splitter which splits the light beam internal to the system into a redirected beam and an output light beam;

a light to frequency converter which receives said redirected light beam from the beam splitter internal to the system and generates a pulse train having a frequency proportional to an intensity of light in said redirected light beam;

a counter that maintains a count of pulses in the pulse train such that the count of pulses provides a measure of the intensity of light in said redirected light beam;

a light intensity circuit which monitors the ongoing count of pulses maintained by the counter and adjusts one or more of the plurality of color channels in real time to thereby control intensity of the output light beam; and

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

20. The system of claim 19, further comprising a host computer which triggers said light engine and triggers said camera.

21. The system of claim 19, wherein the 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.

22. The system of claim 19, 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, Fö

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