Devices and method for enrichment and alteration of cells and other particles

• US 9,174,222 B2
• Filed: 09/14/2011
• Issued: 11/03/2015
• Est. Priority Date: 04/05/2005
• Status: Active Grant

• Current Assignee: Gpb Scientific, LLC, Mehmet Toner
• Original Assignee: Gpb Scientific, LLC, The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Huang, Lotien Richard, Carvalho, Bruce L., Kapur, Ravi, Vernucci, Paul, Toner, Mehmet, Wang, Zihua, Barber, Thomas
• Primary Examiner(s): BROWN, MELANIE YU
• Application Number:
  US13/232,781
• Publication Number:
  US 20120196273A1
• Time in Patent Office: 1,511 Days
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/0472 Diffusion
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• 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 1/30 Combinations with other devices, not otherwise provided for
• CPC Class Codes: B03C 1/32 acting on the medium containing the substance being separated, e.g. magneto-gravimetric-, magnetohydrostatic-, or magnetohydrodynamic separation
• CPC Class Codes: B03C 2201/18 Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: B33Y 80/00 Products made by additive manufacturing
• 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: C12M 47/06 Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 1/40 : Concentrating samples
• CPC Class Codes: G01N 1/4077 : by other techniques involving separation of suspended solids
• CPC Class Codes: G01N 2035/00237 : Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 33/5044 : involving specific cell types
• CPC Class Codes: Y10T 137/0318 : Processes
• CPC Class Codes: Y10T 137/8593 : Systems
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

The invention features devices and methods for the deterministic separation of particles. Exemplary methods include the enrichment of a sample in a desired particle or the alteration of a desired particle in the device. The devices and methods are advantageously employed to enrich for rare cells, e.g., fetal cells, present in a sample, e.g., maternal blood and rare cell components, e.g., fetal cell nuclei. The invention further provides a method for preferentially lysing cells of interest in a sample, e.g., to extract clinical information from a cellular component, e.g., a nucleus, of the cells of interest. In general, the method employs differential lysis between the cells of interest and other cells (e.g., other nucleated cells) in the sample.

Claims

1. A method for transferring particles from a first fluid to a second fluid, wherein said second fluid is different from said first fluid, the method comprising:

(a) applying a first fluid comprising particles to a first channel of a device, wherein said device comprises at least two channels;

(b) flowing said first fluid through said first channel in said device wherein said first channel comprises at least one array of obstacles that, in response to the flow of said first fluid, laterally displaces said particles within said array towards a second channel, wherein each of said arrays comprises a plurality of rows of obstacles wherein each row comprises a plurality of obstacles and is shifted laterally with respect to a previous row by a distance smaller than a distance between obstacles in said previous row, wherein said distance between obstacles in a row is the same in all rows in a given array, and wherein said lateral shift of rows and said distance cause each of said arrays to direct particles having a hydrodynamic size above a critical size and below said distance between obstacles into the array and in a direction towards said second channel;

(c) flowing said second fluid through said second channel in said device and flowing particles laterally displaced in step (b) through said second channel, wherein said second channel is arranged along at least a portion of said at least one array of obstacles in said first channel; and

(d) continuing the flow of particles in step (c) until said particles exit said second channel in said second fluid.

2. The method of claim 1, wherein said particles are cells.

3. The method of claim 2, wherein said cells are lysed, stained or labeled in either said first channel or said second channel.

4. The method of claim 3, wherein said cells are fluorescently labeled.

5. The method of claim 1, wherein said particles are contacted in said first channel, said second channel, or in both said channels, with a reagent that chemically or physically alters the particles.

6. The method of claim 1, wherein particles exiting said second channel in step (d) enter a third channel comprising a third fluid, wherein said third fluid is different from said second fluid.

7. The method of claim 1, wherein said device comprises at least one bypass channel that is devoid of obstacles that laterally displace said particles.

8. The method of claim 1, wherein said particles are laterally displaced into a channel in which said particles react with a reagent and the reacted particles are then moved into a separate channel to remove unreacted reagent or reaction byproducts.

9. The method of claim 1, wherein reagents are added to said particles to increase the hydrodynamic size of said particles.

10. The method of claim 1, wherein said first channel comprises at least two arrays of obstacles, and wherein said critical size of the first array is larger than a critical size of the second array.

11. A method for transferring cells from a blood sample to a second fluid, wherein said second fluid is different from said blood sample, the method comprising:

(a) applying a blood sample comprising cells to a device, wherein said device comprises at least two channels;

(b) flowing said blood sample through a first channel in said device wherein said first channel comprises at least one array of obstacles that, in response to the flow of said blood sample, laterally displaces said cells within said array towards a second channel, wherein each of said arrays comprises a plurality of rows of obstacles wherein each row comprises a plurality of obstacles and is shifted laterally with respect to a previous row by a distance smaller than a distance between obstacles in said previous row, wherein said distance between obstacles is the same in all rows in a given array, and wherein said lateral shift of rows and said distance cause each of said arrays is configured to direct particles having a hydrodynamic size above a critical size and below said distance between obstacles into the array and in a direction towards said second channel;

(c) flowing said second fluid through said second channel in said device and flowing the cells laterally displaced in step (b) through said second channel, wherein said second channel is arranged along at least a portion of said at least one array of obstacles in said first channel; and

(d) continuing the flow of cells in step (c) until said cells exit said second channel in said second fluid.

12. The method of claim 11, wherein said at least one array of obstacles is configured to separate red blood cells from other cells.

13. The method of claim 12, wherein said other cells are contacted in said first channel, in said second channel, or in both said channels with a reagent that chemically or physically alters the cells.

14. The method of claim 13, wherein said other cells are lysed, stained or labeled in either said first channel or said second channel.

15. The method of claim 14, wherein said other cells are fluorescently labeled.

16. The method of claim 11, wherein said array of obstacles is configured to separate white blood cells from red blood cells and platelets in the blood.

17. The method of claim 16, wherein said white blood cells are contacted in said first channel, and/or in said second channel, or in both said channels with a reagent that chemically or physically alters the cells.

18. The method of claim 11, wherein said cells from said blood sample are laterally displaced into a channel in which said cells react with a reagent and the reacted cells are then moved into a separate channel to remove unreacted reagent or reaction byproducts.

19. The method of claim 11, wherein reagents are added to said cells to increase the hydrodynamic size of said cells.