초록 ▼

The invention relates generally to fluid processing and, in particular aspects, processing fluids for detection, selection, trapping and/or sorting of particulate moieties. Sheath flow devices described allow isolation of target species from fluid samples while avoiding non-specific binding of unwan

The invention relates generally to fluid processing and, in particular aspects, processing fluids for detection, selection, trapping and/or sorting of particulate moieties. Sheath flow devices described allow isolation of target species from fluid samples while avoiding non-specific binding of unwanted species to the surfaces of the separation device. Biological fluid processing, detection, sorting or selection of cells, proteins, and nucleic acids is described. The invention finds particular use in diagnostic settings, analyzing a patient's medical condition, monitoring and/or adjusting a therapeutic regimen and producing cell based products.

대표청구항 ▼

1. A sheath flow device comprising: (a) a first laminar flow establishing surface upstream of a sheath flow plane area configured to establish a sheath laminar flow in the sheath flow plane area;(b) a second laminar flow establishing surface parallel to the sheath flow plane area and upstream of a s

1. A sheath flow device comprising: (a) a first laminar flow establishing surface upstream of a sheath flow plane area configured to establish a sheath laminar flow in the sheath flow plane area;(b) a second laminar flow establishing surface parallel to the sheath flow plane area and upstream of a sample sheath flow plane area, wherein the second laminar flow establishing surface is configured to establish a sample laminar flow in the sheath flow plane area; and(c) a particle movement station configured to deflect a target species from the sample laminar flow and into the sheath laminar flow, wherein the particle movement station has a substantially rectangular interior space for bounding the sample laminar flow and the sheath laminar flow, wherein the interior space has first and second lateral dimensions transverse to the sample laminar flow, wherein the second lateral dimension is at least about 2 times larger than the first lateral dimension such that nonspecific binding is reduced along the larger of the first and second lateral dimensions. 2. The sheath flow device of claim 1, further comprising (d) a third laminar flow establishing surface upstream of a second sheath flow plane area. 3. The sheath flow device of claim 2, wherein the laminar flow establishing surfaces in (a) (b) and (d) are sufficient to maintain separate sheath flow planes substantially free of turbulent flow. 4. The sheath flow device of claim 3, further comprising a structural member having a first surface and a second surface, wherein the first surface and the second surface are opposite each other, wherein the first surface comprises the second laminar flow establishing surface of subpart (b), and wherein the second surface comprises the third laminar flow establishing surface of subpart (d). 5. The sheath flow device of claim 4, wherein the particle movement station employs at least one of a magnetic force, an acoustic force, an electrophoretic force and an optical force. 6. The sheath flow device of claim 5, adapted for magnetophoretic particle separation of a target species. 7. The sheath flow device of claim 6, further comprising at least one reservoir containing a reagent. 8. The sheath flow device of claim 6, wherein the target species is a stem cell and the sample laminar flow comprises whole blood. 9. The sheath flow device of claim 6, that is microfluidic. 10. The sheath flow device of claim 6, further comprising a reservoir containing a reagent, said reagent comprising at least one of a buffer, a quantity of magnetic beads, a stem cell expansion agent, an aptamer, a composition comprising a protein, a bacterial cell culture, and a composition comprising a bacteriophage population. 11. A method of separating a target species from a fluid sample in a sheath flow device, comprising: (a) providing the device of claim 1;(b) establishing a laminar sheath of buffer flow along a planar surface of the sheath flow device;(c) establishing a laminar flow of the fluid sample adjacent to the laminar sheath of buffer flow; and(d) deflecting the target species from the laminar flow of the fluid sample and into the laminar sheath of buffer flow; wherein the laminar sheath of buffer flow and the laminar flow of the fluid sample are adjacent. 12. The method of claim 11, further comprising establishing a second laminar sheath of buffer flow, adjacent to the laminar flow of the fluid sample, wherein (b) comprises establishing the laminar flow of the fluid sample between the first and second laminar buffer flows. 13. The method of claim 12, wherein establishing the first and second laminar sheaths of buffer flow and the laminar flow of the fluid sample are sufficient to maintain separate flow planes substantially free of turbulent flow. 14. The method of claim 13, wherein deflecting the target species from the laminar flow of the fluid sample and into the laminar sheath of buffer flow comprises using at least one of a magnetic force, an acoustic force, an electrophoretic force and an optical force. 15. The method of claim 14, wherein the target species comprises at least one of a cell, a bacterium, a virus, a protein and a nucleic acid. 16. The method of claim 15, wherein the targets species is isolated in a trapping station over which the laminar sheath of buffer flow passes. 17. The method of claim 16, wherein the trapping station employs magnetic force in order to trap the targets species selectively labeled with magnetic particles. 18. The method of claim 17, wherein the target species is a circulating tumor cell and the fluid sample is whole blood. 19. A method of analyzing a patient the method comprising: (a) providing the flow device as recited in claim 1;(b) receiving a sample comprising tumor cells from a patient;(c) separating said tumor cells from the sample by a separation method comprising: (i) labeling the sample with magnetic particles having a specific affinity for said tumor cells, thereby producing a labeled sample, (ii) passing the labeled sample through the fluidic device comprising a sorting region having a magnetic field gradient effective to deflect and/or trap the magnetic particles from the labeled sample and thereby separate the tumor cells from the sample, wherein the sample flows in a sheath of buffer solution to reduce nonspecific binding of the sample to the fluidic device; and(d) characterizing the tumor cells separated from the sample in (c). 20. The method of claim 19, wherein the characterizing in (c) provides information for diagnosing a condition, screening for a clinical trial, assessing the effectiveness of a therapeutic treatment, and measuring the effectiveness of surgery. 21. The method of claim 19, wherein the sample is a fluid sample taken from the patient. 22. The method of claim 19, wherein the sample is not taken from a biopsy of the patient. 23. The method of claim 19, wherein the sample is a blood sample. 24. The method of claim 23, wherein the tumor cells are circulating tumor cells from a non-haematologic cancer. 25. The method of claim 19, wherein the characterizing is a count of said tumor cells. 26. The method of claim 19, wherein the characterizing is a molecular characterization of said tumor cells. 27. The method of claim 26, wherein said molecular characterization is a genetic mutation in said tumor cells. 28. A method of monitoring and, if appropriate, adjusting a patient's treatment regimen, the method comprising: (a) providing the flow device as recited in claim 1;(b) receiving a sample comprising tumor cells from a patient undergoing a first treatment regimen;(c) separating said tumor cells from the sample by a separation method comprising: (i) labeling the sample with magnetic particles having a specific affinity for said tumor cells, thereby producing a labeled sample, (ii) passing the labeled sample through the fluidic device comprising a sorting region having a magnetic field gradient effective to deflect and/or trap the magnetic particles from the labeled sample and thereby separate the tumor cells from the sample, wherein the sample flows in a sheath of buffer solution to reduce nonspecific binding of the sample to the fluidic device; and(d) characterizing the tumor cells separated from the sample in (c) to suggest a future treatment for the patient. 29. The method of claim 28, wherein the first treatment regimen is a chemotherapy regimen. 30. The method of claim 29, wherein the future treatment is a different chemotherapy regimen. 31. The method of claim 28, wherein suggesting the future treatment for the patient comprises predicting a future effectiveness of the first treatment regimen. 32. The method of claim 28, wherein suggesting a future treatment for the patient comprises identifying a second treatment regimen that is different than the first treatment regimen and accounts for a characteristic of the tumor cells not previously observed for the patient. 33. The method of claim 32, further comprising: (d) receiving a sample comprising tumor cells from the patient after the patient has undergone the second treatment regimen; and(e) thereafter performing (b) and (c) on the sample and tumor cells received in (d). 34. A method of providing cell based products, the method comprising: (a) providing the flow device as recited in claim 1;(b) receiving a sample comprising target cells;(c) separating said target cells from the sample by a separation method comprising: (i) labeling the target cells with magnetic particles having a specific affinity for said target cells, thereby producing a population of labeled cells in the sample, (ii) passing the sample through the fluidic device comprising a sorting region having a magnetic field gradient effective to deflect and/or trap at least a portion of the population of labeled cells from the sample and thereby separate the target cells from the sample, wherein the sample flows in a sheath of buffer solution to reduce nonspecific binding of the sample to the fluidic device; and(d) deriving a cell based product from the target cells separated from the sample in (c). 35. The method of claim 34, further comprising treating the target cells separated from the sample in (b) to produce the cell based product. 36. The method of claim 35, wherein the target cells are stem cells and treating the target cells comprising treating the stem cells to become an effective therapeutic agent. 37. The method of claim 36, wherein treating the stem cells to become an effective therapeutic agent comprises differentiating the stem cells to produce a more specific cell type. 38. The method of claim 34, wherein the sample is a fluid sample taken from a patient. 39. The method of claim, 38, wherein the fluid sample is a blood sample. 40. The method of claim 34, further comprising characterizing the target cells separated from the sample in (b). 41. The method of claim 40, wherein the characterizing is a count of said target cells. 42. The method of claim 40, wherein the characterizing is a molecular characterization of said target cells. 43. The method of claim 42, wherein said molecular characterization is a genetic sequence of said target cells. 44. A fluidic separating device comprising: (a) at least one sample inlet channel configured to provide a sample stream in the fluidic separating device;(b) at least one sheath flow inlet channel configured to provide one or more fluid sheaths within the fluidic separating device and separating the sample stream from a surface of the device, thereby reducing nonspecific binding of components of the sample to the device; and(c) a sorting station fluidly coupled to the sample and sheath flow inlets and located along a path of the sample stream, wherein the sorting station has a substantially rectangular interior space for bounding the one or more fluid sheaths, wherein the interior space has first and second lateral dimensions transverse to the sample stream's direction of flow, wherein the second lateral dimension is at least about 2 times larger than the first lateral dimension such that nonspecific binding is reduced along the larger of the first and second lateral dimensions, and wherein during operation the sorting station has a magnetic field gradient effective to deflect and/or trap magnetic particles from the sample stream. 45. The fluidic separating device of claim 44, wherein the at least one sheath flow inlet channel is configured to provide two sheath streams separated from one another by said sample stream along the first lateral dimension. 46. The fluidic separating device of claim 44, wherein the sorting station has a substantially rectangular interior space for bounding fluid flowing through the sorting station, and wherein the interior space is defined, in part, by two substantially parallel and substantially planar surfaces separated by a distance not greater than about 2 millimeters, andwherein the at least one sheath flow inlet channel is configured to provide two sheath streams flowing in contact with the two parallel planar surfaces and separated from one another by said sample stream. 47. The fluidic separating device of claim 46, wherein the two substantially parallel and substantially planar surfaces are separated by a distance not greater than about 1 millimeter. 48. The fluidic separating device of claim 46, wherein the at least one sheath flow inlet channel comprises a first sheath flow inlet channel, which comprises a substantially planar surface located upstream of the sorting station and oriented substantially parallel to the two substantially planar surfaces of the interior space. 49. The fluidic separating device of claim 48, further comprising a second sheath flow inlet channel, which comprises its own substantially planar surface located upstream of the sorting station and oriented substantially parallel to the two substantially planar surfaces of the interior space. 50. The fluidic separating device of claim 49, wherein the at least one sample inlet channel comprises its own substantially planar surfaces oriented substantially parallel to the two substantially planar surfaces of the interior space.