Focused acoustics for detection and sorting of fluid volumes

• US 6,849,423 B2
• Filed: 12/28/2001
• Issued: 02/01/2005
• Est. Priority Date: 11/29/2000
• Status: Expired due to Fees

• Current Assignee: Labcyte, Inc. (Danaher Corporation)
• Original Assignee: Picoliter Inc. (Danaher Corporation)
• Inventors: Ellson, Richard N., Mutz, Mitchell W., Lee, David Soong-Hua
• Primary Examiner(s): GITOMER, RALPH J
• Application Number:
  US10/040,926
• Publication Number:
  US 20020090720A1
• Time in Patent Office: 1,131 Days
• Field of Search: 435/29, 435/173.1, 435/173.9, 435/325, 365/200
• US Class Current: 435/29
• CPC Class Codes: B01D 2015/389 using ultra-sound
• CPC Class Codes: B41J 2/14008 Structure of acoustic ink jet print heads
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: G01N 15/1023 Microstructural devices for non-optical measurement
• CPC Class Codes: G01N 2015/1028 Sorting particles
• CPC Class Codes: G01N 2015/1415 Control of particle position
• CPC Class Codes: G01N 2015/142 Acoustic or ultrasonic focussing
• CPC Class Codes: G01N 2035/1039 Micropipettes, e.g. microcapillary tubes
• CPC Class Codes: G01N 29/0681 by acoustic microscopy, e.g. scanning acoustic microscopy
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 35/1074 arranged in a two-dimensional array

Abstract

A method is provided for acoustically ejecting from a channel or other container a plurality of fluid droplets, each of which contains one or more particles or other localized volumes. The localized volumes, which can be living cells, are ejected towards sites on a substrate surface, a container, or a channel. An integrated cell sorting and arraying system is also provided that is capable of sorting based upon cellular properties by the selective ejection of cells from a carrier fluid. The cells can be ejected with adjustable velocity and trajectory. The ejected cells can be directed to form an array, wherein each site of the array can contain a single cell. Additionally provided is a method of forming arrays of single live cells more efficiently, rapidly, flexibly, and economically than by other cell array approaches. This method permits efficient, continuous, and simultaneous sorting of cells based upon the quantitative or semiquantitative measurement of cellular properties, and also permits non-binary or severally-branched decision-making. An integrated system, which includes a processor, and methods are also provided for the detection, selection, and ejection of selected particles or circumscribed volumes, such as live cells, from a continuous stream of fluid-suspended particles or other circumscribed volumes flowing in channels.

Claims

1. A separation method comprising the steps of:

(a) detecting, in a fluid having a surface and containing a plurality of localized fluid volumes having a different acoustic impedance than the fluid, a single localized volume located sufficiently near the surface for ejection;

(b) determining whether the single localized volume possesses one or more properties;

(c) selecting the single localized volume for ejection from the fluid based on the determination of one or more properties in step (b); and

(d) ejecting the single localized volume from the fluid towards a suitable substrate material by use of focused radiation.

2. The method of claim 1, wherein the focused radiation is focused acoustic radiation.

3. The method of claim 2, wherein the acoustic radiation is delivered to the localized volume at a geometric center of the volume.

4. The method of claim 3, wherein the geometric center of the volume is located approximately 50 to 150 μ

m beneath the fluid surface.

5. The method of claim 1, wherein the focused radiation is focused electromagnetic radiation.

6. The method of claim 1, wherein the localized volume comprises a solid or gel particle.

7. The method of claim 1, wherein the localized volume comprises a cell.

8. The method of claim 7, wherein the localized volume comprises a living cell.

9. The method of claim 1, wherein the localized volume is ejected in a trajectory substantially perpendicular to the fluid surface.

10. The method of claim 1, wherein the localized volume is ejected with a first non-zero velocity component perpendicular to the fluid surface and a second non-zero velocity component parallel to the fluid surface to effect a nonvertical trajectory whereby the localized volume experiences a net displacement in a direction parallel to the fluid surface.

11. The method of claim 10, wherein the nonvertical trajectory is directionally controllable.

12. The method of claim 10, wherein the non-vertical distance of travel parallel to the fluid surface is controllable by varying the focused energy.

13. The method of claim 10, wherein the vertical distance of travel parallel to the fluid surface is controllable by varying the focused energy.

14. The method of claim 1, wherein the determining in step (b) of the one or more properties is a quantitative or semiquantitative determination.

15. The method of claim 14, wherein the fluid is contained in a fluidic channel on a substrate surface.

16. The method of claim 15, wherein the acoustic impedances of step 1(a) and the properties of step (b) are inputted into a processor as individual parameters and further wherein upon selection of an acoustic impedance and one or more properties as inputted in the processor, the selection and ejection of the localized volume of steps (c) and (d), respectively, is carried out according to the inputted parameters.

17. The method of claim 15, wherein the fluidic channel has a cross section permitting the localized volume to flow freely through the channel.

18. The method of claim 17, wherein the cross section is sufficiently narrow such that if two or more of the localized volumes are present in the channel, they necessarily form a linear flow path of single localized volumes successively passing any given point in the channel.

19. The method of claim 17, wherein the fluid is contained in two or more fluidic channels on a substrate surface.

20. The method of claim 1, wherein the localized volumes are circumscribed fluid volumes.

21. The method of claim 20, wherein the circumscribed volumes are comprised of cells such that a single individual cell comprises a single circumscribed volume.

22. The method of claim 21, wherein the cells are living cells.

23. The method of claim 22, wherein the fluid is contained in a fluidic channel on a substrate surface.

24. The method of claim 23, wherein the fluidic channel has a cross section permitting each cell to flow freely through the channel.

25. The method of claim 24, wherein the cross section is sufficiently narrow such that if two or more cells are present in the channel, they necessarily form a linear flow path of single cells successively passing any given point in the channel.

26. The method of claim 22, wherein steps (a) through (d) are repeated so as to selectively eject a plurality of living cells from the fluid.

27. The method of claim 26, wherein each cell is ejected toward and deposited on a different target site on a substrate surface.

28. The method of claim 27, wherein the target site at which each cell is deposited is selected based on the determination of the one or more properties in step (b).

29. The method of claim 26, wherein each cell is ejected toward and deposited into a target container.

30. The method of claim 29, wherein cells having a first property as determined in step (b) are deposited into a first target container, and cells having a second property as determined in step (b) are deposited into a second target container.

31. The method of claim 30, wherein additional cells having a particular property as determined in step (b) are deposited into an additional target container.

32. The method of claim 29, wherein the target container is a well in a well plate.

33. The method of claim 29, wherein the target container is a fluidic channel in a substrate surface.

34. The method of claim 31, wherein the target containers are individual wells in a single well plate.

35. The method of claim 31, wherein the target containers are individual fluidic channels in a substrate surface.

36. The method of claim 1, wherein the fluid is contained in two or more fluidic channels on a substrate surface.

37. The method of claim 1, wherein steps (a) through (d) are repeated so as to eject a plurality of single localized volumes from the fluid.