Systems and method for correction of positionally dependent electromagnetic radiation detected from objects within a fluid column

Patent metadata

Field	Value
Publication number	US-11858832-B2
Application number	US-202318099137-A
Country	US
Kind code	B2
Filing date	Jan 19, 2023
Priority date	Mar 16, 2020
Publication date	Jan 2, 2024
Grant date	Jan 2, 2024

Abstract

A discrimination system that forms a fluid column and interrogates objects within the fluid column with an excitation source. An optical arrangement collects output electromagnetic radiation emanating from the excited objects disposed within the fluid column and directs the output electromagnetic radiation to a detector. An analyzer reduces the positional dependency of the detected intensity by normalizing the value based on the position of each object.

First claim

We claim:

1. A discrimination system, comprising:
an excitation source that generates excitation electromagnetic radiation directed toward cells at a measurement region, where the cells emanate output electromagnetic radiation in response to the excitation electromagnetic radiation;
collection optics having an optical axis, wherein the collection optics collect the output electromagnetic radiation from the cells;
at least one detector that generates an electrical signal responsive to the intensity of the output electromagnetic radiation collected by the collection optics, wherein the at least one detector generates a signal representative of a cell's position passing through the measurement region relative to the optical axis of the collection optics; and
an analyzer having instructions stored thereon i) to normalize the intensity of the output electromagnetic radiation represented in the electrical signal by adjusting the detected intensity of the output electromagnetic radiation based on the position of the cells in the measurement region relative to the optical axis of the collection optics, and ii) to discriminate a first type of cell from other cells with the adjusted output electromagnetic radiation intensity.

2. The discriminating system of claim 1 , wherein the at least one detector comprises a first detector and a second detector that detects the position of cells in the measurement region.

3. The discriminating system of claim 2 , wherein the second detector comprises a position detector.

4. The discriminating system of claim 3 , wherein the position detector is one selected from the group of: a camera, a CCD, PSD, SiPM split detector, and photodiode array.

5. The discrimination system of claim 1 , wherein the at least one detector comprises a first detector that: i) generates an electrical signal responsive to the intensity of the output electromagnetic radiation collected by the collection optics and ii) detects the position of cells in the measurement region.

6. The discrimination system of claim 5 , wherein the first detector is not located in an image plane of the collection optics.

7. The discrimination system of claim 5 , wherein the first detector comprises one selected from the group consisting of a: PMT split detector, SiPM split detector, and photodiode array, an array of PMTs, and an array of SiPMs.

8. The discrimination system of claim 1 , wherein the cells are located in a generally elliptical cross section of a core stream contained within a generally circular cross section of sheath fluid, and wherein cells in the measurement region are located at differing positions on a major axis of the generally elliptical cross section of the core stream.

9. The discrimination system of claim 8 , wherein the instructions stored on the analyzer for normalizing the intensity of the output electromagnetic radiation represented in the electrical signal based on the position of the cells in the measurement region applies a correction to the intensity of the output electromagnetic radiation represented in the electrical signal.

10. The discrimination system of claim 9 , wherein the amount of correction is determined for a cell at each position on the major axis of the core stream, and wherein the correction is applied to each event based on the position of the cell associated with the event on the major axis of the core stream.

11. The discriminating system of claim 1 , wherein the first type of cell comprises one selected from the group consisting of: viable X-chromosome bearing sperm; and viable Y-chromosome bearing sperm.

12. The discriminating system of claim 1 , wherein the analyzer includes instructions stored thereon for discriminating viable X-chromosome bearing sperm and viable Y-chromosome bearing sperm.

13. The discriminating system of claim 1 , wherein the cell comprises sperm located within an elliptical column of sample fluid, where the sample fluid is formed coaxially with an outer layer of sheath fluid and the sheath fluid comprises a generally cylindrical shape.

14. The discriminating system of claim 1 , further comprising an element that modifies the output electromagnetic radiation to increase the uniformity of output electromagnetic radiation collected by the collection optics of cells at different positions.

15. A detection system, comprising:
collection optics having an optical axis, wherein the collection optics collect output electromagnetic radiation from cells at a measurement region;
at least one detector that generates an electrical signal responsive to the intensity of the output electromagnetic radiation collected by the collection optics, wherein the at least one detector generates a signal representative of a cell's position passing through the measurement region relative to the optical axis of the collection optics; and

an analyzer having instructions stored thereon i) to normalize the intensity of the output electromagnetic radiation represented in the electrical signal by adjusting the detected intensity of the output electromagnetic radiation based on the position of the cells in the measurement region relative to the optical axis of the collection optics, and ii) to discriminate a first type of cell from other cells with the adjusted output electromagnetic radiation intensity.

16. The detection system of claim 15 , wherein the at least one detector comprises a first detector and a second detector, where a mathematical operation on the electrical signal provided by the first detector is related to the position of cells in the measurement region based on a signal from the second detector.

17. The detection system of claim 16 , wherein the second detector comprises a position detector.

18. The detection system of claim 17 , wherein the position detector is one selected from the group consisting of: a camera, a CCD, PSD, SiPM split detector, and photodiode array.

19. The detection system of claim 15 , wherein the at least one detector comprises a first detector that: i) generates an electrical signal responsive to the intensity of the output electromagnetic radiation collected by the collection optics and ii) detects the position of cells in the measurement region.

20. The detection system of claim 19 , wherein the first detector comprises one selected from the group consisting of a: PMT split detector, SiPM split detector, and photodiode array, an array of PMTs, and an array of SiPMs.

21. The detection system of claim 19 , wherein the first detector is located outside an image plane of collection optics.

22. The detection system of claim 15 , wherein the cells are located in a generally elliptical cross section of a core stream contained within a generally circular cross section of sheath fluid and where cells are located at differing positions on a major axis of the generally elliptical cross section of core stream, wherein the instructions stored on the analyzer for normalizing the intensity of the output electromagnetic radiation represented in the electrical signal based on the position of the cells in the measurement region applies a correction to the intensity of the output electromagnetic radiation represented in the electrical signal.

23. The detection system of claim 22 , wherein the correction is determined for each position on the major axis of the core stream, and wherein the correction is applied to each event based on the position of the cell associated with the event on the major axis of the core stream.

Assignees

• Inguran Llc

Inventors

• Kiesel Peter
• Karin Todd
• Evans Kenneth Michael

Classifications

• C02F1/325 Primary

  Irradiation devices or lamp constructions · CPC title

• G01N15/14

  Optical investigation techniques, e.g. flow cytometry · CPC title

• G01N15/1404

  Handling flow, e.g. hydrodynamic focusing · CPC title

• G01N15/1429 Primary

  Signal processing · CPC title

• G01N15/1434 Primary

  Optical arrangements · CPC title