Apparatus and method for continuous particle separation

• US 7,735,652 B2
• Filed: 05/31/2007
• Issued: 06/15/2010
• Est. Priority Date: 06/01/2006
• Status: Active Grant

• Current Assignee: The Trustees of Princeton University (Princeton University)
• Original Assignee: The Trustees of Princeton University (Princeton University)
• Inventors: Davis, John, Austin, Robert, Inglis, David William, Sturm, James
• Primary Examiner(s): Mackey; Patrick H
• Assistant Examiner(s): MATTHEWS, TERRELL HOWARD
• Application Number:
  US11/809,559
• Publication Number:
  US 20080023399A1
• Time in Patent Office: 1,111 Days
• Field of Search: 209/155, 209/208, 204/451, 210/767
• US Class Current: 209/155
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2400/0472 Diffusion
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• 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: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2001/4072 membraneless transfer of a component between two parallel laminar flows of fluid
• CPC Class Codes: G01N 2015/012 Red blood cells
• CPC Class Codes: G01N 2015/016 White blood cells
• CPC Class Codes: G01N 2015/018 Platelets
• CPC Class Codes: G01N 2035/00465 Separating and mixing arrangements
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 35/025 having a carousel or turntable for reaction cells or cuvettes

Abstract

The invention is directed to an apparatus and a method of separating particles, such as cells, from a heterogeneous fluid, such as blood, where the particles have a large range of sizes.

Claims

1. A continuous flow fluidic particle separation device, comprising:

a sample input;

a carrier fluid input;

a plurality of separation areas in fluid connection with the sample input, each separation area configured and adapted to separate particles having a size equal to or greater than a critical size from particles having a size less than the critical size in a flow of a heterogeneous fluid sample through each separation area, the flow having a flow direction, wherein the critical size for each separation area is different, and wherein particles having a size less than the critical size for a separation area and particles having at least the critical size for the separation area move in different directions in the separation area, the particles having a size less than the critical size moving with the flow direction of the fluid sample, and the particles having at least the critical size moving at an angle to the flow direction of the fluid sample;

at least one non-clogging pathway in fluid connection with the carrier fluid input, configured and adapted to accept separated particles from at least one of the separation areas into a flow of the carrier fluid in the non-clogging pathway;

a sample output in fluid connection with the separation areas; and

at least one alternate output in fluid connection with the non-clogging pathway, wherein

when a heterogeneous fluid sample containing particles to be separated by size is introduced into the device through the sample input, and a carrier fluid is introduced into the device through the carrier fluid input;

the fluid sample passes through the separation areas;

the carrier fluid passes through the non-clogging pathway in contact with the fluid sample in the separation areas;

no mixing other than diffusion occurs between the liquids; and

particles having at least the critical size separated in each of the separation areas pass into the carrier fluid.

2. The device of claim 1, wherein particles in the heterogeneous fluid sample have a size range of from less than 1 μ

m to greater than 10 μ

m.

3. The device of claim 1, wherein particles in the heterogeneous fluid sample have a size range of from less than 1 μ

m to greater than 20 μ

m.

4. The device of claim 1, wherein separated particles that pass into the carrier fluid have a size at least that of the critical size.

5. The device of claim 1, further comprising a non-clogging pathway and an alternate output for each separation area, wherein each output has a fluidic resistance that is substantially the same as the other outputs.

6. The device of claim 1, wherein at least one of the separation areas comprises an array comprising a network of gaps within a microfluidic channel.

7. The device of claim 6, wherein the array is an ordered array of obstacles.

8. The device of claim 6, wherein the array is an ordered array of micro- or nano-fabricated features.

9. The device of claim 1, wherein the device has a flow rate of at least about 1000 μ

m/second.

10. The device of claim 1, wherein the device has a throughput of at least about 1 μ

l/minute.

11. A method of separating particles by size in a continuous fluid flow, the method comprising:

introducing a heterogeneous fluid sample into a sample input of a continuous flow fluidic particle separation device, and introducing a carrier fluid into a carrier fluid input of the device, thereby providing a sample flow through a plurality of separation areas and a carrier flow parallel to and in contact with the sample flow, wherein each of the flows has a sufficiently low Reynolds number that there is substantially no turbulence, and mixing of the fluids in the flow occurs only through diffusion;

separating particles having a size at least that of a predetermined critical size from particles having a size less than the critical size, wherein each separation area has a different critical size, and wherein particles having a size less than the critical size move with the sample flow, and particles having a size at least that of the critical size move at an angle to the sample flow; and

passing the separated particles into the carrier fluid.

12. The method of claim 11, wherein the fluid sample is blood, and the particles comprise blood cells.

13. The method of claim 11, further comprising separating the particles having a size at least that of the critical size.

14. The method of claim 11, further comprising removing particles separated from a plurality of the separation areas through a single output.

15. The method of claim 11, further comprising removing particles separated in each of the separation areas through separate outputs.

16. The method of claim 11, further comprising removing all particles having a size of at least about 1 μ

m from the sample fluid in the separation areas.

17. The method of claim 16, wherein the carrier fluid comprises phosphate buffered saline.

18. The method of claim 16, wherein the carrier fluid comprises at least one of an additive that prevents adhesion and anticoagulant.