Multiple laminar flow-based particle and cellular separation with laser steering

• US 20100216208A1
• Filed: 03/02/2010
• Published: 08/26/2010
• Est. Priority Date: 07/31/2002
• Status: Active Grant

• Current Assignee: ABS Global, Inc. (Genus PLC)
• Original Assignee: Arryx, Inc. (Haemonetics Corporation)
• Inventors: Anderson, Amy, Rosenbaum, Neil Harris, Shireman, Jessica, Plewa, Joseph, Mueth, Daniel, Gruber, Lewis
• Granted Patent: US 8,158,927 B2
• Time in Patent Office: Days
• US Class Current: 435/173.900
• CPC Class Codes: A61L 2/08 Radiation
• CPC Class Codes: B01D 21/0009 Settling tanks making use of electricity or magnetism electric ultra filters B01D61/425; filters making use of electricity or magnetism B01D35/06; magnetic or electrostatic separation B03C
• CPC Class Codes: B01D 21/01 using flocculating agents for purifying water C02F1/52; for liquid radioactive waste G21F9/10
• CPC Class Codes: B01D 21/262 by using a centrifuge
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0454 radiation pressure, optical tweezers
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B03C 1/00 Magnetic separation
• CPC Class Codes: B03C 1/002 High gradient magnetic separation acting directly on the substance being separated B03C1/025; acting on the medium containing the substance being separated B03C1/32
• CPC Class Codes: B03C 1/288 disposed at the outer circumference of a recipient
• CPC Class Codes: B03C 2201/26 for use in medical or biological applications
• CPC Class Codes: B03C 5/005 Dielectrophoresis, i.e. dielectric particles migrating towards the region of highest field strength
• CPC Class Codes: B04B 13/00 Control arrangements specially designed for centrifuges; Programme control of centrifuges control arrangements for feed, charge, or discharge B04B11/00
• CPC Class Codes: G01N 15/0205 by optical means
• CPC Class Codes: G01N 15/0227 using imaging; using holography
• CPC Class Codes: G01N 15/04 Investigating sedimentation of particle suspensions
• CPC Class Codes: G01N 15/042 by centrifuging and investigating centrifugates centrifuges per se B04B
• CPC Class Codes: G01N 15/05 in blood
• CPC Class Codes: G01N 15/0631 : Separation of liquids, e.g. by absorption, wicking
• CPC Class Codes: G01N 15/0656 : using electric, e.g. electrostatic methods or magnetic methods by investigating individual particles G01N15/1031, G01N15/12
• CPC Class Codes: G01N 15/14 : Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: G01N 15/149 : specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/0053 : in liquids, e.g. trouble
• CPC Class Codes: G01N 2015/0233 : using holography
• CPC Class Codes: G01N 2015/0288 : Sorting the particles
• CPC Class Codes: G01N 2015/035 : the optical arrangement forming an integrated apparatus with the sample container
• CPC Class Codes: G01N 2015/045 : by optical analysis
• CPC Class Codes: G01N 2015/1016 : Particle flow simulating, e.g. liquid crystal cell
• CPC Class Codes: G01N 2015/1028 : Sorting particles
• CPC Class Codes: G01N 21/6486 : Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 2201/061 : Sources
• CPC Class Codes: G01N 2458/00 : Labels used in chemical analysis of biological material
• CPC Class Codes: G01N 27/447 : using electrophoresis
• CPC Class Codes: G01N 27/44756 : Apparatus specially adapted therefor
• CPC Class Codes: G01N 33/487 : of liquid biological material
• CPC Class Codes: G01N 33/491 : by separating the blood components G01N15/05 takes precedence
• CPC Class Codes: G01N 33/4915 : using flow cells flow cytometry G01N15/14
• CPC Class Codes: G01N 33/5091 : for testing the pathological state of an organism
• CPC Class Codes: G02B 21/32 : Micromanipulators structurally combined with microscopes
• CPC Class Codes: G03H 1/0005 : Adaptation of holography to specific applications holographic optical element G02B5/32; holographic scanner G02B26/106; recognition using holographic mask G06V10/88; holographic memories G11B7/0065, G11C13/042
• CPC Class Codes: G03H 1/08 : Synthesising holograms, i.e. holograms synthesized from objects or objects from hologramsusing electric digital computers G06F; G06T
• CPC Class Codes: G03H 1/2294 : Addressing the hologram to an active spatial light modulator
• CPC Class Codes: G03H 2001/0077 : for optical manipulation, e.g. holographic optical tweezers [HOT]
• CPC Class Codes: G03H 2001/085 : Kinoform, i.e. phase only encoding wherein the computed field is processed into a distribution of phase differences
• CPC Class Codes: G03H 2225/32 : Phase only
• CPC Class Codes: G21K 5/08 : Holders for targets or for other objects to be irradiated
• CPC Class Codes: H05H 3/04 : Acceleration by electromagnetic wave pressure

Abstract

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.

Claims

1. (canceled)

2. A method of sorting particles in a microfluidic channel, comprising:

providing a microfluidics chip having at least one microfluidic channel, said microfluidics channel containing particles in a solution;

providing a holographic optical trapping apparatus including a laser emitting a laser beam, and a diffractive optical element to diffract said laser beam into a plurality of beamlets, said plurality of beamlets being used to produce a plurality of optical traps to trap said particles in said solution;

encoding a surface of said diffractive optical element with a sequence of phase patterns in order to implement a corresponding sequence of holographic optical trapping patterns; and

trapping and arranging said particles in a plurality of parallel lines in said microfluidic channel, wherein a spacing between said parallel lines permits said trapped particles to be moved in predetermined directions specified by said arrangement of said plurality of parallel lines of said optical traps.

3. The method according to claim 2, further comprising:

timing an extinction one of said parallel lines of said optical traps on either side of at least one of said particles, such that said at least one particle is moved in a forward or a reverse direction.

4. The method according to claim 3, further comprising:

moving said optical traps to a predetermined area.

5. The method according to claim 4, further comprising:

collecting said optical traps from said predetermined area.

6. The method according to claim 3, further comprising:

reducing said spacing between said parallel lines of said optical traps, or varying a curvature of said parallel lines of said optical traps, such that said particles are disposed in a focusing pattern to concentrate said particles.

7. The method according to claim 3, further comprising:

increasing said spacing between said parallel lines of said optical traps, or varying a curvature of said parallel lines of said optical traps, such that said particles are dispersed.

8. The method according to claim 3, further comprising:

increasing said spacing between said parallel lines of said optical traps, or narrowing said spacing between said parallel lines of said optical traps, to speed up or slow down movement of said particles, respectively.

9. The method according to claim 2, wherein said diffractive optical element is mounted on a prism, and each of said sequence of phase patterns is rotated into place by a motor.

10. The method according to claim 2, wherein said diffractive optical element is placed on a perimeter of a disk, and said disk is rotated by a motor to cycle through said sequence of phase patterns.

11. The method according to claim 2, further comprising:

providing a rectilinear array of optical traps within said solution;

wherein said particles are one of trapped when a trapping force of said optical traps exceeds an external bias force of fluid flow of said solution, or flow past said rectilinear array when said bias force exceeds said trapping force.

12. The method according to claim 11, wherein when said bias force exceeds said trapping force to a differing degree for different fractions of each of said particles, said particles hop from one of said optical traps to another of said optical traps along a direction of a principal axis of said rectilinear array.

13. The method according to claim 11, wherein said particles are deflected to greater angles based on said trapping force of said rectilinear array or a relatively increased size of said particles, than said particles affected by said fluid flow of said solution and a relatively decreased size of said particles.

14. The method according to claim 13, further comprising:

orienting said rectilinear array to achieve an angle of optimal deflection of said particles; and

adjusting an intensity of said optical gradient forces exerted on said particles such that said particles of said relatively increased size hop from said one of said optical traps to said another of said optical traps along said direction of said principal axis of said rectilinear array.

15. The method according to claim 14, further comprising:

adding additional rectilinear arrays downstream of said rectilinear array.

56. The method according to claim 2, wherein said particles are cells.

57. The method according to claim 56, wherein the cells are sperm cells.

16. An apparatus for sorting particles, comprising:

a microfluidics chip having at least one microfluidic channel, said microfluidics channel containing a solution with particles therein;

a holographic optical trapping apparatus including;

a laser which emits a laser beam;

a diffractive optical element which diffracts said laser beam into a plurality of beamlets, said plurality of beamlets being used to produce a plurality of optical traps to trap said particles in said solution;

wherein a surface of said diffractive optical element is encoded with a sequence of phase patterns in order to implement a corresponding sequence of holographic optical trapping patterns; and

wherein said particles are trapped and arranged by said holographic optical trapping apparatus in a plurality of parallel lines in said microfluidic channel, such that a spacing between said parallel lines permits said trapped particles to be moved in predetermined directions specified by said arrangement of said plurality of parallel lines of said optical traps.

17. The apparatus according to claim 16, further comprising:

a prism on which said diffractive optical element is mounted; and

a motor which rotates each of said sequence of phase patterns on said surface of said diffracted optical element into place.

18. The apparatus according to claim 16, further comprising:

a disk on which said diffractive optical element is placed at a perimeter thereof; and

a motor which rotates said disk to cycle through said sequence of phase patterns.

19. The apparatus according to claim 16, further comprising:

a rectilinear array of optical traps provided within said solution;

wherein said particles are one of trapped when a trapping force of said optical traps exceeds an external bias force of fluid flow of said solution, or flow past said rectilinear array when said bias force exceeds said trapping force.

20. The apparatus according to claim 19, wherein when said bias force exceeds said trapping force to a differing degree for different fractions of each of said particles, said particles hop from one of said optical traps to another of said optical traps along a direction of a principal axis of said rectilinear array.

21. The apparatus according to claim 20, wherein said rectilinear array is oriented to achieve an angle of optimal deflection of said particles;

and

wherein an intensity of said optical gradient forces exerted on said particles is adjusted such that said particles of said relatively increased size hop from said one of said optical traps to said another of said optical traps along a direction of a principal axis of said rectilinear array.

22. The apparatus according to claim 21, further comprising:

adding additional rectilinear arrays downstream of said rectilinear array.

58. The apparatus according to claim 16, wherein the particles are cells.

59. The apparatus according to claim 58, wherein the cells are sperm cells.

23. An apparatus for sorting particles, comprising:

an intake through which a sample of particles is introduced;

a distribution disk rotated by motor control;

a sample delivery system for providing said sample of said particles onto said distribution disc;

a holographic optical trapping system for imaging and optically trapping said particles; and

a control and analysis system which provides information for said holographic optical trapping system to use in sorting said particles.

24. The apparatus according to claim 23, wherein said particles are cells.

25. The apparatus according to claim 23, further comprising:

a plurality of chambers for collecting said particles.

26. The apparatus according to claim 25, further comprising:

means for distributing and holding said particles on said distribution disk such that they can be measured.

27. The apparatus according to claim 26, wherein said distributing and holding means includes one of fluid chambers, gels, adhesives, waxes, or freezing mechanisms.

60. The apparatus according to claim 24, wherein the cells are sperm cells.

28. A method of sorting particles, comprising:

introducing a sample of particles into a spinning disc-based sorter through an intake thereof;

delivering said particles onto a distribution disk which is rotated by motor control;

distributing said particles over said distribution disk;

holding said particles on said distribution disk;

imaging said particles using a holographic optical trapping system;

using a control and analysis system to determine which particles to sort; and

removing wanted or unwanted of said particles from said holding means on said distribution disk by optically trapping said particles.

29. The method according to claim 28, wherein said particles are cells.

30. The method according to claim 28, further comprising:

collecting said wanted or unwanted particles in a respective chamber.

31. The method according to claim 28, wherein said determining step includes measuring said particles using said holographic optical trapping apparatus.

32. The method according to claim 28, wherein said holding step includes one of housing said particles in fluid chambers, immobilizing said particles using gels, binding said particles using adhesives or waxes, or freezing said particles into a solid mass.

61. The method according to claim 29, wherein the cells are sperm cells.

33. A method of purifying and sorting particles, comprising:

growing particles in high concentrations in a chamber;

flowing a culture medium through said chamber to collect products from said particles, said products being provided in a waste media;

flowing said waste media from said chamber into a sorter through a first channel;

flowing a solution of beads coated with predetermined affinity sites into said sorter through a second channel;

mixing said waste media and said bead solution in a first mixer to provide a first mixture in which said products bind to said affinity sites on said beads;

flowing said first mixture through a first separation region along with a first input buffer from a third channel;

separating said first mixture into beads with said bound products and waste in said first separation region, said beads with said bound products being flowed into a fourth channel using passive diffusive sorting;

discarding said waste from said first separation region into a first output channel;

flowing said beads with said bound products from said fourth channel into a second mixer along with a denaturing solution from a fifth channel;

mixing said beads with said bound products in said second mixer with said denaturing solution from said fifth channel, to break bonds at said affinity sites between said products and said beads, to form a second mixture of purified products separated from said beads;

flowing said second mixture from said second mixer, through a second output channel along with a second input buffer solution from a sixth channel, into a second separation region;

discarding said beads through a third output channel by passive diffusive sorting in sad second separation region; and

removing said purified products from said second separation region through a seventh channel.

34. The method according to claim 33, wherein said particles are cells.

62. The method according to claim 34, wherein the cells are sperm cells.

35. An apparatus for purifying and sorting particles, comprising:

a chamber in which particles are gown at high concentrations, and a culture medium is flowed through to collect products from said particles, said products being provided in a waste media;

a sorter, comprising;

a first channel though which said waste media is flowed into said sorter from said chamber;

a second channel through which a solution of beads coated with predetermined affinity sites is flowed into said sorter;

a first mixer for mixing into a first mixture, said waste media from said first channel and said solution of beads from said second channel, wherein said products bind to said affinity sites on said beads;

a third channel through which a first input buffer is flowed into a first separation region along with said first mixture from said first mixer;

a fourth channel into which said beads with bound products from said first separation region are separated by passive diffusive sorting from waste from said first mixture;

a first output channel into which said waste from said first mixture is discarded;

a fifth channel through which a denaturing solution is flowed into a second mixer along with said beads with said bound products from said fourth channel, to form a second mixture of said products separated from said beads;

a second output channel into which said products separated from said beads is flowed into a second separation region to be purified;

a sixth channel through which a second input buffer solution is flowed into said second separation region where said products are purified;

a third output channel through which said beads are discarded by passive diffusive sorting; and

a seventh channel through which said purified products are removed from said sorter.

36. The apparatus according to claim 35, wherein said particles are cells.

63. The apparatus according to claim 36, wherein the cells are sperm cells.

37. An apparatus from sorting particles, comprising:

a flow cell having a fluid input channel for introducing a sample of particles in a laminar flow, said particles being disposed in a plurality of input lines having a predetermined separation therebetween to determine a width of said fluid input channel;

two output channels having a same width as said fluid input channel, said two output channels being disposed on either side of said fluid input channel, each having a buffer solution in a laminar flow flowing therethrough;

a sorting region having no mechanical separation between said fluid input channel and said two output channels and said laminar flows therein;

an optical trapping system which produces an array of a plurality of funneling optical traps in said fluid input channel proximate to said sorting region, said optical traps which separate said particles in said fluid input channel by a minimum predetermined distance;

wherein said optical trapping system includes a relatively high-numerical-aperture objective lens used to implement said array of optical traps; and

a detection system which measures said particles to determine which particles are sorted from said sorting region of said fluid input channel into said two output channels.

38. The apparatus according to claim 37, wherein said optical traps are a pattern of relatively low intensity optical traps established by a set of static holograms mounted on a rotating wheel, such that said pattern changes as a function of a pattern of rotation.

39. The apparatus according to claim 37, wherein said optical traps located at a farthest downstream position of said fluid input channel are of fixed intensity and position, said optical traps which maintain a separation between said input lines of flow.

40. The apparatus according to claim 37, wherein said optical traps located at an upstream position of said fluid input channel are of changeable intensity and position, in order to disturb a flow of particles when said particles are clumped, and to pass through said particles when said particles are individual or unclumped particles.

41. The apparatus according to claim 37, wherein said detection system is one of a fluorescence detection system, a scattering measurement system, or an optical deflection system.

42. The apparatus according to claim 37, wherein a field-of-view of said objective lens is a same as a width of each of said fluid input channel and two output channels.

43. The apparatus according to claim 37, wherein said optical trapping system includes spatial light modulators that create phase masks to drive said optical trapping system.

44. The apparatus according to claim 43, wherein update rates of said spatial light modulators are 30 Hz or more.

45. The apparatus according to claim 37, wherein said particles are cells.

46. A method of sorting particles, comprising:

introducing a sample of particles into a laminar flow in a fluid input channel of a flow cell, said particles being disposed in a plurality of input lines having a predetermined separation therebetween to determine a width of said fluid input channel;

introducing a buffer solution into a laminar flow into two output channels having a same width as said fluid input channel, said two output channels being disposed on either side of said fluid input channel;

providing a sorting region having no mechanical separation between said fluid input channel and said two output channels and said laminar flows therein;

producing an array of a plurality of funneling optical traps in said fluid input channel proximate to said sorting region, using an optical trapping system, said optical traps which separate said particles in said fluid input channel by a minimum predetermined distance;

wherein said optical trapping system includes a relatively high-numerical-aperture objective lens used to implement said array of optical traps; and

measuring said particles using a detection system to determine which particles are sorted from said sorting region of said fluid input channel into said two output channels.

47. The method according to claim 46, wherein said optical traps are a pattern of relatively low intensity optical traps established by a set of static holograms mounted on a rotating wheel, such that said pattern changes as a function of a pattern of rotation.

48. The method according to claim 46, wherein said optical traps located at a farthest downstream position of said fluid input channel are of fixed intensity and position, said optical traps which maintain a separation between said input lines of flow.

49. The method according to claim 46, wherein said optical traps located at an upstream position of said fluid input channel are of changeable intensity and position, in order to disturb a flow of particles when said particles are clumped, and to pass through said particles when said particles are individual or unclumped particles.

50. The method according to claim 46, wherein said detection system is one of a fluorescence detection system, a scattering measurement system, or an optical deflection system.

51. The method according to claim 46, wherein a field-of-view of said objective lens is a same as a width of each of said fluid input channel and two output channels.

52. The method according to claim 46, wherein said optical trapping system includes spatial light modulators that create phase masks to drive said optical trapping system.

53. The method according to claim 52, wherein update rates of said spatial light modulators are 30 Hz or more.

54. The method according to claim 46, wherein said particles are cells.

55. The method according to claim 54, wherein the cells are sperm cells.