Multiple laminar flow-based particle and cellular separation with laser steering
The invention provides a method, apparatus and system for separating blood and other types of cellular components, and can be combined with holographic optical trapping manipulation or other forms of optical tweezing. One of the exemplary methods includes providing a first flow having a plurality of blood components; providing a second flow; contacting the first flow with the second flow to provide a first separation region; and differentially sedimenting a first blood cellular component of the plurality of blood components into the second flow while concurrently maintaining a second blood cellular component of the plurality of blood components in the first flow. The second flow having the first blood cellular component is then differentially removed from the first flow having the second blood cellular component. Holographic optical traps may also be utilized in conjunction with the various flows to move selected components from one flow to another, as part of or in addition to a separation stage.
1. A method of particle washing to remove undesirable component comprising
providing a first flow having a plurality of particles;
providing a second flow;
identifying the particle to be selected;
killing or damaging the particle creating undesirable component or additional undesirable component;
contacting the first flow with the second flow to provide at least a first separation region;
washing the undesirable component from the first flow to the second flow at the first separation region;
maintaining the selected particle in the first flow; and
collecting the first flow.
2. The method of claim 1 , wherein killing or damaging the particle comprises killing or damaging an unselected particle.
3. The method of claim 1 , wherein maintaining the particle in the first flow comprises maintain a selected particle.
4. The method of claim 1 , further comprising
contacting the first flow with the second flow to provide at least an additional separation region;
washing the undesirable component from the first flow to the second flow at the additional separation region; and
maintaining the particle in the first flow; wherein the particle is a selected particle.
5. The method of claim 1 , wherein the particle is a cell.
6. The method of claim 5 , wherein the cell is a sperm cell.
7. The method of claim 1 , wherein the undesirable component is selected from the group consisting of debris; a dead cell or a damaged cell; a non-sperm cell; a bacteria, a virus, a yeast, and a combination thereof.
8. The method of claim 7 , the debris is cellular debris.
9. The method of claim 1 , wherein the killing or damaging the selected particles creating undesirable components or additional undesirable component comprises using light energy from a laser.
10. The method of claim 1 , wherein the washing the undesirable component from the first flow to second flow comprises utilizing differential sedimentation rates between the undesirable component and the particle.
11. The method of claim 1 , wherein the washing the undesirable component from the first flow to second flow comprises utilizing differential densities between the undesirable component and the particle.
12. The method of claim 1 , wherein the washing the undesirable component from the first flow to second flow comprises utilizing differential diffusions between the undesirable component and the particle.
13. The method of claim 1 , wherein the washing the undesirable component from the first flow to second flow comprises utilizing differential motilities between the undesirable component and the particle.
14. The method of claim 1 , wherein the washing the undesirable component from the first flow to second flow comprises utilizing optical force targeting to separate the undesirable component and the particle.
15. The method of claim 1 , wherein the washing the undesirable component from the first flow to second flow comprises utilizing dielectrophoresis to separate the undesirable component and the particle.
16. The method of claim 1 , wherein the washing the undesirable component from the first flow to second flow comprises utilizing electrophoresis to separate the undesirable component and the particle.
17. The method of claim 1 , wherein the washing the undesirable component from the first flow to second flow comprises utilizing magnetic force to separate the undesirable component and the particle.
18. The method of claim 1 , wherein the washing the undesirable component from the first flow to second flow comprises utilizing differential surface tension or differential surface chemistry to separate the undesirable component and the particle.
19. The method of claim 1 , wherein the first flow and the second flow are substantially laminar and substantially non-turbulent.
20. The method of claim 1 , wherein the separation region has a predetermined length substantially parallel to a direction of flow of the first and second flows and a predetermined depth substantially perpendicular to the direction of flow of the first and second flows, the predetermined length and predetermined depth having been determined from a first sedimentation rate of the undesirable component, from a second sedimentation rate of the particle, from a first flow rate of the first flow, and from a second flow rate of the second flow.
21. The method of claim 1 , wherein the first flow and the second flow have substantially the same flow rates.
22. The method of claim 1 , wherein the first flow has a first flow rate and the second flow has a second flow rate, wherein the second flow rate is comparatively greater than the first flow rate.
23. The method of claim 1 , wherein identifying the particle comprises using spectroscopic methods and using a computer program to analyze the spectral data to identify the selected particles.
24. The method of claim 1 , wherein identifying the particle comprises distinguishing cells bearing an X or Y chromosome.
25. The method of claim 1 , further comprising
providing a third flow;
contacting the first flow with the third flow to provide a second separation region;
washing the undesirable component from the first flow to the third flow at the second separation region; and
maintaining the particle in the first flow; wherein the particle is a selected particle.
26. The method of claim 25 , wherein the second flow and the third flow constrict the first flow, particularly orthogonally to the first flow at the separation region.
27. The method of claim 25 , wherein the plurality of particles is aligned such that the plurality of particles is oriented parallel to the confronting wall and is flattened such that the plurality of particles is in a singular file, particularly at the separation region.
28. A method of particle washing comprising
providing a first flow having a plurality of particles;
providing a second flow;
identifying the particle to be selected;
killing or damaging the particle creating undesirable component or additional undesirable component;
contacting the first flow with the second flow to provide a first separation region;
washing the particle from the first flow to the second flow at the first separation region;
maintaining the undesirable component in the first flow; and
collecting the second flow.
29. The method of claim 28 , wherein killing or damaging the particle comprises killing or damaging an unselected particle.
30. The method of claim 28 , wherein washing the particle comprises washing a selected particle.
31. The method of claim 28 , further comprising
contacting the first flow with the second flow to provide at least an additional separation region;
washing the particle from the first flow to the second flow at the additional separation region; wherein the particle is a selected particle; and
maintaining the undesirable component in the first flow.
32. The method of claim 28 , wherein the particle is a cell; and wherein the undesirable component is selected from the group consisting of debris; a dead cell or a damaged cell; a non-sperm cell; a bacteria, a virus, a yeast, and a combination thereof.
33. The method of claim 32 , wherein the cell is a sperm cell; and wherein the debris is cellular debris.
34. The method of claim 32 , wherein the washing the particle from the first flow to second flow comprises utilizing differential sedimentation rates between the undesirable component and the particle.
35. The method of claim 28 , wherein the washing the particle from the first flow to second flow comprises utilizing a technique selected from the group consisting of separating by density, separating by diffusivity, separating by motility, separating by optical forces, separating by using dielectrophoresis, separating by using electrophoresis, separating by using magnetic forces, separating by using surface tension or surface chemistry, and a combination thereof.
36. The method of claim 28 , wherein identifying the particle comprises using spectroscopic methods and using a computer program to analyze the spectral data to identify the particle.
37. The method of claim 28 , wherein identifying the particle comprises distinguishing cell bearing an X or Y chromosome.
38. The method of claim 28 , further comprising
providing a third flow;
contacting the first flow with the third flow to provide a second separation region;
washing the particle from the first flow to the third flow at the second separation region;
wherein the particle is a selected particle; and
maintaining the undesirable component in the first flow.
39. The method of claim 38 , wherein the second flow and the third flow constrict the first flow, particularly orthogonally to the first flow at the separation region;
wherein the plurality of particles is aligned such that the plurality of particles is oriented parallel to the confronting wall and flattened such that the plurality of particles is in a singular file, particularly at the separation region.
40. An apparatus for washing undesirable component from particle comprising
at least a first channel having a first inlet for a first flow and a second inlet for a second flow;
the first channel further having a first outlet for the first flow and a second outlet for the second flow;
wherein the first flow comprises a plurality of particles;
an identifying apparatus that identifies the particle to be selected in the channel;
a killing apparatus that kills or damages the unselected particle in the channel;
wherein killing or damaging the unselected particle creates undesirable component or additional undesirable component; and
wherein the first channel is adapted to allow the undesirable component in the first flow to move differentially into the second flow while maintaining the selected particle in the first flow or to allow the selected particle in the first flow to move differentially into the second flow while maintaining the undesirable component in the first flow.
41. The apparatus of claim 40 , further comprising
at least a second channel having a first inlet for a first flow and second inlet for a second flow;
the second channel further having a first outlet for the first flow and a second outlet for the second flow; and
wherein the second channel is adapted to allow the undesirable component in the first flow to move differentially into the second flow while maintaining the selected particle in the first flow or to allow the selected particle in the first flow to move differentially into the second flow while maintaining the undesirable component in the first flow.
42. The apparatus of claim 41 , wherein the second channel has a third inlet and a third outlet for a third flow; and
wherein the second channel is adapted to allow the undesirable component in the first flow to move differentially into the third flow while maintaining the selected particle in the first flow or to allow the selected particle in the first flow to move differentially into the third flow while maintaining the undesirable component in the first flow.
43. The apparatus of claim 40 , wherein the particle is a cell; and, wherein the undesirable component is selected from the group consisting of debris; a dead cell or a damaged cell; a non-sperm cell; a bacteria, a virus, a yeast, and a combination thereof.
44. The apparatus of claim 43 , wherein the cell is a sperm cell; and wherein the debris is cellular debris.
45. The apparatus of claim 40 , wherein the identifying apparatus uses spectroscopic methods and uses a computer program to analyze the spectral data to identify the particle.
46. The apparatus of claim 40 , wherein the identifying apparatus can distinguish cells bearing an X or Y chromosome.
47. The apparatus of claim 40 , wherein the killing apparatus comprises a laser.
48. The apparatus of claim 40 , wherein the first channel has a third inlet and a third outlet for a third flow; and
wherein the first channel is adapted to allow the undesirable component in the first flow to move differentially into the third flow while maintaining the selected particle in the first flow or to allow the selected particle in the first flow to move differentially into the third flow while maintaining the undesirable component in the first flow.
49. The apparatus of claim 48 or claim 42 , wherein the channels adapted to allow the undesirable component or the selected particle in the first flow to move differentially into the second flow and the third flow utilizes differential sedimentation rates between the undesirable component and the particle.
50. The apparatus of claim 48 or claim 42 , wherein the channels adapted to allow the undesirable component or the selected particle in the first flow to move differentially into the second flow and the third flow utilizes a technique selected from the group consisting of separating by density, separating by diffusivity, separating by motility, separating by optical forces, separating by using dielectrophoresis, separating by using electrophoresis, separating by using electrophoresis, separating by using magnetic forces, separating by using surface tension or surface chemistry, and a combination thereof.
51. The apparatus of claim 48 or claim 42 , wherein the first, second and third flows are substantially laminar and substantially non-turbulent; and
wherein the second flow and the third flow constrict the first flow, particularly orthogonally to the first flow at the channel.
52. The apparatus of claim 48 or claim 42 , wherein the plurality of particles is aligned such that the plurality of particles is oriented parallel to the confronting wall and flattened such that the plurality of particles is in a singular file in the channel.