Light collection from objects within a fluid column
An optical arrangement receives output light emanating from an object disposed within a fluid column that crosses an optical refraction boundary of the fluid column between the object and the optical arrangement. The optical arrangement modifies the output light such that the modified output light has an intensity that is more uniform than the output light.
1. An optical apparatus, comprising:
an optical arrangement configured to receive output light emanating from an object disposed within a fluid column, the output light crossing an optical refraction boundary of the fluid column between the object and the optical arrangement, the optical arrangement configured to modify the output light such that the modified output light has an intensity that is more uniform than the output light by preferentially attenuating light received by the optical arrangement from a first position in the fluid column compared to light received from a second position in the fluid column, wherein the second position is farther from the center of the fluid column than the first position along an axis perpendicular to the optical axis of the optical arrangement; and
a detector configured to detect the modified output light and to provide an electrical signal responsive to the modified output light.
2. The optical apparatus of claim 1 , wherein the intensity of the modified output light is substantially uniform across a cross section of the fluid column.
3. The optical apparatus of claim 1 , further comprising one or more of:
an excitation light source arranged to provide excitation light to the object within the fluid column, the object emanating the output light in response to the excitation light;
an optical filter configured to substantially block the excitation light from reaching the detector; and
a focusing lens arranged to collect the modified output light and to focus the modified output light toward the detector.
4. The optical apparatus of claim 1 , further comprising electrical circuitry configured to receive the electrical signal from the detector and to distinguish between an object of a first type and an object of a second type based on the electrical signal.
5. The optical apparatus of claim 4 , wherein:
an amplitude of the electrical signal is responsive to an intensity of the modified output light; and
the electrical circuitry is configured to distinguish between the first type of object and the second type of object by comparing the amplitude of the electrical signal to a threshold value.
6. An optical arrangement configured to receive output light emanating from an object disposed within a fluid column, the output light crossing an optical refraction boundary between the object and the optical arrangement, the optical arrangement configured to modify the output light such that the modified output light has an intensity that is substantially uniform irrespective of a position of the object,
wherein the optical arrangement is configured to preferentially attenuate light received by the optical arrangement from a first position in the fluid column compared to light received from a second position in the fluid column, wherein the second position is farther from the center of the fluid column than the first position along an axis perpendicular to the optical axis of the optical arrangement.
7. The optical arrangement of claim 6 , wherein the optical arrangement comprises a mask configured to attenuate light from the first position in the fluid column.
8. The optical arrangement of claim 7 , wherein the mask comprises a wire.
9. The optical arrangement of claim 6 , wherein the optical arrangement is configured to preferentially collect a first portion of the output light emanating from the object at larger angles with respect to the optical axis relative to a second portion of the output light emanating from the object at smaller angles with respect to the optical axis.
10. The optical arrangement of claim 9 , wherein the optical arrangement comprises a split objective lens configured to preferentially collect the first portion of the output light.
11. The optical arrangement of claim 10 , wherein:
the first portion of light includes:
output light emanating from the object at positive angles with respect to the optical axis of the optical arrangement; and
output light emanating from the object at negative angles with respect to the optical axis of the optical arrangement; and
the split objective lens comprises:
a first lens configured to receive the light emanating from the object at positive angles with respect to the optical axis of the optical arrangement; and
a second lens configured to receive the light emanating from the object at negative angles with respect to the optical axis of the optical arrangement.
12. The optical arrangement of claim 8 , wherein the optical arrangement further includes a mask configured to attenuate the second portion of the output light.
13. A discrimination system, comprising:
an excitation light source configured to generate excitation light and to direct the excitation light toward an object in a fluid column, the object emanating output light in response to the excitation light;
an optical arrangement configured to receive the output light, the output light crossing an optical refraction boundary of the fluid column between the object and the optical arrangement, the optical arrangement configured to modify the output light such that the modified output light has an intensity that is more uniform than the output light, wherein the optical arrangement comprises an elongated mask feature that attenuates the output light received from the first position, a length of the elongated mask feature being much greater than a width of the elongated mask feature, the elongated mask feature disposed proximate to an active area of the detector such that a longitudinal axis of the elongated mask feature is substantially parallel to a direction of flow of the objects in the fluid column;
an optical detector configured to detect the modified output light and to provide an electrical signal responsive to the modified output light; and
object type discrimination circuitry configured to discriminate between a first type of object and a second type of object based on the electrical signal.
14. The system of claim 13 , wherein the intensity of the modified output light is substantially uniform irrespective of a position of the object.
15. The system of claim 13 , wherein the circuitry is configured to perform X/Y sperm cell discrimination.
16. The system of claim 13 , wherein the elongated mask feature has a round cross sectional area.
17. The system of claim 13 , wherein the elongated mask feature has a rectangular cross sectional area.
18. The system of claim 13 , wherein the elongated mask feature is a metal wire.
19. The system of claim 13 , wherein the elongated mask feature is a trace deposited on glass.
20. The system of claim 13 , wherein the elongated mask feature is a filament.
21. The system of claim 13 , wherein the elongated mask feature is an ink line printed on transparent paper.
22. The system of claim 13 , wherein the elongated mask feature is disposed at about a center of an active area of the detector.
23. A method, comprising:
receiving, by an optical apparatus, output light from an object in a fluid column that has crossed an optical refraction boundary of the fluid column;
modifying the output light such that the modified output light has an intensity that is substantially uniform irrespective of a position of the object in the fluid column by attenuating a first portion of the output light received from a first position in the fluid column relative to a second portion of output light received from a second position in the fluid column, wherein the second position is farther from the center of the fluid column than the first position along an axis perpendicular to the optical axis of the optical apparatus;
detecting the modified output light; and
generating an electrical signal in response to the modified output light.
24. The method of claim 23 , wherein modifying the output light further comprises:
preferentially collecting a first portion of the output light emanating from the object at higher angles with respect to the optical axis relative to collecting a second portion of the output light emanating from the object at lower angles with respect to the optical axis.
25. A flow cytometer system, comprising:
an excitation light source configured to generate excitation light and to direct the excitation light toward an object in a fluid column, the object emanating output light in response to the excitation light;
an optical arrangement configured to receive the output light, the output light crossing an optical refraction boundary of the fluid column between the object and the optical arrangement, the optical arrangement configured to modify the output light such that the modified output light has an intensity that is more uniform than an intensity of the output light, the optical arrangement comprising a plate that includes an aperture and an elongated mask feature disposed at least partially across the aperture, the elongated mask feature arranged such that a longitudinal axis of the elongated mask feature is substantially parallel to a direction of flow of the objects in the fluid column, the plate and the elongated mask feature being less transmissive to the output light than the aperture;
an optical detector disposed adjacent to the plate and arranged to receive the modified output light transmitted through the aperture, the optical detector configured to detect the modified output light and to provide an electrical signal responsive to the modified output light; and
object type discrimination circuitry configured to discriminate between a first type of object and a second type of object based on the electrical signal.
26. The system of claim 25 , wherein the plate and the elongated mask feature are one unitary structure.
27. The system of claim 25 , wherein the elongated mask feature is attached to the plate.
28. The system of claim 25 , wherein optical transparency of the plate to the output light varies as a function of position across the plate.
29. The system of claim 25 , wherein opposing longitudinal edges of the elongated mask feature are substantially parallel to one another.
30. The system of claim 25 , wherein opposing longitudinal edges of the elongated mask feature are not parallel to one another.
31. The system of claim 25 , wherein opposing longitudinal edges of the elongated mask feature are tapered along a longitudinal axis of the elongated mask feature.