Method and apparatus for sorting cells
Apparatus for sorting and orienting sperm cells has a pair or walls in confronting relationship forming a flow chamber having inlet, a downstream outlet, and intermediate detector region. The inlet receives first and second spaced apart streams of input fluid and a third stream of sample fluid containing the cells to be sorted. The first and second streams have respective flow rates relative to third stream, such that the third stream is constricted forming a relatively narrow sample stream, so that the cells are oriented parallel to the walls. A detector detects desired cells and a sorter downstream of the detector for sorting the desired cells from the stream.
1. A sensor for imaging an object in an object plane comprising:
an optical source for illuminating the object using k-vector imaging;
a collection lens provided a predetermined distance from the object plane equal to an effective focal length of the collection lens;
wherein the illumination of the object produces beams which exit the collection lens and are collimated as beamlets which are divided to form multiple images of the object in the form of light leaving the object plane at different angles relative to a central axis of the sensor; and
wherein a lateral positioning of each collimated beamlet is determined primarily by its angle from the object plane; and
a detector responsive to light from the object.
2. The apparatus of claim 1 wherein the detector is responsive to at least one of light directed along the optical axis of the sensor and light directed off the axis of the detector.
3. The apparatus of claim 2 wherein the off-axis detector comprises at least two photo detectors laterally displaced with respect to the central axis for detecting off-axis light corresponding to different positions in the object plane.
4. The apparatus of claim 3 further including a filter for filtering at least one of fluorescent and scattered light from each beam.
5. The apparatus of claim 3 wherein the photo detector comprises a multiple element photo multiplier tube for detecting multiple beams of light.
6. The apparatus of claim 1 wherein the optical source comprises a laser.
7. The apparatus of claim 6 further including a lens for collimating the light from each object into separate beams.
8. The apparatus of claim 7 further including collecting lens for each beam.
9. The apparatus of claim 7 further including a splitter for splitting on-axis light from off-axis light.
10. The apparatus of claim 1 further including a cell killer downstream of the detector for killing undesired cells.
11. The apparatus of claim 10 where the cell killer comprises at least one of a laser and electrode for directing lethal energy at the undesired cells.
12. The apparatus of claim 10 wherein the cell killer comprises a source of light for activating a lethal target in the cell.
13. The apparatus of claim 1 further including a cell electroporation for effectively processing or killing undesired cells.
14. The apparatus of claim 1 wherein the flow device is operable at about 2° C.-10° C. to prolong cell life.
15. A sensor for imaging an object in an object plane comprising:
an optical sensor using k-vector imaging;
a beamsplitter that breaks an input beam from the optical source into multiple beams;
a first collimating lens that collimates the multiple beams from the beamsplitter into parallel beams;
wherein the parallel beams are focused onto the object in a sample stream in the object plane or onto multiple objects in multiple streams in the object plane;
a collection lens provided a predetermined distance from the object plane equal to an effective focal length of the collection lens;
wherein an illumination of the object produces beams which exit the collection lens and are collimated as beamlets which are divided to form multiple images of the object in the form of light leaving the object plane at different angles relative to a central axis of the sensor; and
wherein a lateral positioning of each collimated beamlet is determined primarily by its angle from the object plane and
a detector responsive to light from the object.
16. The apparatus of claim 15, wherein the detector is responsive to at least one of light directed along the optical axis of the sensor and light directed off the axis of the detector.
17. The apparatus of claim 16, wherein the off-axis detector comprises at least two photo detectors laterally displaced with respect to the central axis for detecting off-axis light corresponding to different positions in the object plane.
18. The apparatus of claim 17 further including a filter for filtering at least one of fluorescent and scattered light from each beam.
19. The apparatus of claim 17 wherein the photo detector comprises a multiple element photo multiplier tube for detecting multiple beams of light.
20. The apparatus of claim 15 wherein the optical source comprises a laser.
21. The apparatus of claim 20 further including a lens for collimating the light from each object into separate beams.
22. The apparatus of claim 21 further including collecting lens for each beam.
23. The apparatus of claim 21 further including a splitter for splitting on-axis light from off-axis light.
24. The apparatus of claim 15 further including a cell killer downstream of the detector for killing undesired cells.
25. The apparatus of claim 24 where the cell killer comprises at least a laser or at least an electrode for directing lethal energy at the undesired cells.
26. The apparatus of claim 24 wherein the cell killer comprises a source of light for activating a lethal target in the cell.
27. The apparatus of claim 15 further including a cell electroporation for effectively processing or killing undesired cells.
28. The apparatus of claim 15 wherein the flow device is operable at about 2° C.-10° C. to prolong cell life.
29. A sensor for imaging an object in an object plane comprising:
an optical sensor using k-vector imaging;
a collection lens provided a predetermined distance from the object plane equal to an effective focal length of the collection lens;
wherein the object is in a sample stream between at least two sheath streams and wherein the sample stream and the at least two sheath streams have respective flow rate or pressures in a flow chamber such that the sample stream is constricted in a detection region of the flow chamber whereby the object is oriented in a selected direction relative to a beam from the optical source;
wherein an illumination of the object produces beams which exit the collection lens and are collimated as beamlets which are divided to form multiple images of the object in the form of light leaving the object plane at different angles relative to a central axis of the sensor; and
wherein a lateral positioning of each collimated beamlet is determined primarily by its angle from the object plane and
a detector responsive to light from the object.
30. The apparatus of claim 29, wherein the sheath streams flow at a relatively faster flow rate compared to the flow rate of the sample stream, whereby a pressure is exerted from the sheath streams towards the sample stream.
31. The apparatus of claim 29, wherein the sheath streams flow at a relatively slower flow rate compared to the flow rate of the sample stream, whereby a pressure is exerted from the sheath streams towards the sample stream.
32. A sensor for imaging a cell in an object plane comprising:
an optical sensor using k-vector imaging;
a collection lens provided a predetermined distance from the object plane equal to an effective focal length of the collection lens;
wherein an illumination of the cell produces beams which exit the collection lens and are collimated as beamlets which are divided to form multiple images of the object in the form of light leaving the object plane at different angles relative to a central axis of the sensor; and
wherein a lateral positioning of each collimated beamlet is determined primarily by its angle from the object plane;
a detector responsive to light from the object; and
a cell killer downstream of the detector for killing or damaging undesired cells.
33. The apparatus of claim 32 wherein the cell killer comprises at least a laser or at least a cell electroporation for directing lethal or damaging energy at the undesired cells for effectively processing or killing undesired cells.