Systems and methods for in situ spectroscopic measurements

• US 7,319,522 B2
• Filed: 05/27/2005
• Issued: 01/15/2008
• Est. Priority Date: 05/27/2004
• Status: Active Grant

• Current Assignee: Digimarc Corporation, Finesse Solutions, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Finesse Solutions LLC
• Inventors: Havard, John M., Williams, Paul C.
• Primary Examiner(s): Lauchman; Layla G.
• Assistant Examiner(s): UNDERWOOD, JARREAS C
• Application Number:
  US11/139,720
• Publication Number:
  US 20050264817A1
• Time in Patent Office: 963 Days
• Field of Search: 73863-865, 73/84, 356216-236, 356432-448
• US Class Current: 356/436
• CPC Class Codes: G01N 21/0303 Optical path conditioning in cuvettes, e.g. windows; adapted optical elements or systems; path modifying or adjustment G01N21/031 - G01N21/15 take precedence
• CPC Class Codes: G01N 21/532 with measurement of scattering and transmission
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 21/8507 Probe photometers, i.e. with optical measuring part dipped into fluid sample

Abstract

A circularizated semiconductor laser diode (CSLD), such as for example a vertical cavity surface emitting laser (VCSEL) may be used for optical measurements. The CSLD may be used in a cell density probe to perform cell density determination and/or turbidity determination, such as in a biotech, fermentation, or other optical absorbance application. The cell density probe may comprise a probe tip section made from a polytetrafluoroethylene material, which provides sealability, ease of manufacture, durability, cleanability, optical semi-transparency at visible and near infrared wavelengths, and other advantages. The probe tip advantageously provides an optical gap that allows for in situ measurements of optical measurements including but not limited to absorbance, scattering, and fluorescence.

Claims

1. An apparatus for measuring optical absorbance, comprising:

i) a tip section comprising a first light source and a first detector, wherein the first light source emits a first light beam at a first wavelength across a first optical gap towards the first detector, such that the first detector receives the first light beam after the first light beam passes through the first optical gap and generates a first signal representative of the light received at the first detector;

ii) an aperture positioned between the first light source and the first optical gap which aperture substantially prevent light from the first light source from reaching the first detector via a path through that portion of the tip section that is external to the first optical gap; and

iii) a processor coupled to the first detector to determine an absorbance based on the first signal.

2. The apparatus of claim 1, wherein the first light source comprises a CSLD which is a VCSEL, a laser diode having a circularizing lens mounted on one of its facets, or a laser diode having a circularizing lens inside its package.

3. The apparatus of claim 1, wherein the first light source emits at a wavelength in the range of approximately 200 nm to 2000 nm.

4. The apparatus of claim 1 wherein the first light source emits at a near-infrared wavelength.

5. The apparatus of claim 1, further comprising a second detector to detect a characteristic of the first light beam prior to the first light beam being passed through the first optical gap.

6. The apparatus of claim 1, wherein the first and/or the second detectors comprise photodiodes.

7. The apparatus of claim 1, wherein that portion of the tip material that is external to the optical gap comprises polytetrafluoroethylene.

8. The apparatus of claim 1, wherein the tip section further comprises:

a second light source and a second detector, wherein the second light source emits a second light beam at a second wavelength across a second optical gap towards the second detector, such that the second detector receives the second light beam after the second light beam passes through the second optical gap and generates a second signal representative of the light received at the second detector, such that the second signal is also coupled to the processor.

9. The apparatus of claim 1, further comprising a machine-readable medium coupled to the processor and having software stored thereon that can cooperate with the processor to determine the absorbance.

10. The apparatus of claim 1, further comprising electronic circuitry means for supporting operation of the apparatus.

11. The apparatus of claim 1, wherein the determined absorbance is related to cell density.

12. The apparatus of claim 1, further comprising either one or both a wired interface and a wireless interface coupled to the processor to allow communications with the apparatus.

13. The apparatus of claim 1, further comprising means for verifying proper operation of at least some elements of the apparatus.

14. The apparatus of claim 1, further comprising a second light source transmitting a second light beam parallel to the first beam and through the first optical gap onto a second photo-detector, the second light source emitting at a different wavelength than the first light source.

15. The apparatus of claim 14, wherein the first light source emits at a near-infrared wavelength and the second light source emits at an ultraviolet wavelength.

16. The apparatus of claim 14, wherein the first light source emits at a near-infrared wavelength and the second light source emits at a visible wavelength.

17. The apparatus of claim 14, wherein the first light source emits at a near-UV wavelength and the second light source emits at a visible wavelength.

18. An apparatus for making fluorescent measurements comprising:

i) a tip section comprising a first light source and a first detector, wherein the first light source emits a first light beam at a first wavelength across a first optical gap which first beam is incident on a sample present in said first optical gap, such that the first detector receives fluorescent emissions occurring in the sample as a result of the incident light;

ii) an aperture positioned between the first light source and the first optical gap which aperture substantially prevent light from the first light source from reaching the first detector via a path through that portion of the tip section that is external to the first optical gap; and

iii) a processor coupled to the first detector to analyze the fluorescent emissions occurring in the sample as a result of the incident light.

19. An apparatus in accordance with claim 18 wherein said first detector is positioned off the axis of said incident first beam.

20. An apparatus in accordance with claim 18, wherein said light source is a CSLD and said apparatus further comprises:

means for controlling the CSLD;

means for initializing the photometric apparatus;

means for controlling the photometric apparatus;

means for communicating with the photometric apparatus; and

a second light source and second detector.

22. An apparatus in accordance with claim 18, wherein said light source is a CSLD, and said apparatus further comprises:

means for controlling the CSLD;

means for initializing the photometric apparatus;

means for controlling the photometric apparatus;

means for communicating with the photometric device; and

a second light source and second detector.

23. An apparatus in accordance with claim 22 wherein said first detector is positioned off the axis of said incident first beam.

21. An apparatus for making scattering measurements comprising:

i) a tip section comprising a first light source and a first detector, wherein the first light source emits a first light beam at a first wavelength across a first optical gap which first beam is incident on a sample present in said first optical gap, such that the first detector receives light scattered by the sample as a result of the incident light;

ii) an aperture positioned between the first light source and the first optical gap which aperture substantially prevent light from the first light source from reaching the first detector via a path through that portion of the tip section that is external to the first optical gap; and

iii) a processor coupled to the first detector to analyze the light scattered by the sample as a result of the incident light.