Granted Patent B2

US 12,590,955 · App. 15/329,753 · Granted Mar 31, 2026

Inventors: Michael Grisham (Richmond, VA); Curt I. Civin (Baltimore, MD); James C. Sturm (Princeton, NJ); Robert H. Austin (Princeton, NJ); Joseph D'Silva (Princeton, NJ); Yu Chen (Princeton, NJ)

Assignees: ZEON CORPORATION; THE TRUSTEES OF PRINCETON UNIVERSITY; UNIVERSITY OF MARYLAND, BALTIMORE

G01N33/5094B01L3/502746B01L3/502753G01N33/4833B01L2300/0883B01L2400/086

Abstract

Claims (26)

1 . A system for processing first particles from a sample, the first particles comprising leukocytes or stem cells, the system comprising:

a) a microfluidic device, wherein the microfluidic device comprises:

i) a fluidic channel extending from a plurality of inlets to a plurality of outlets;

ii) a first array of obstacles arranged in rows in the channel, wherein each subsequent row of obstacles is shifted laterally with respect to a previous row, wherein the array of obstacles is configured to differentially deflect first particles of at least a predetermined size to a first outlet and second particles of less than the predetermined size to a second outlet, and wherein:

aa) surfaces of two adjacent obstacles in a row of the array of obstacles define a gap;

bb) the two adjacent obstacles defining the gap each have a polygonal cross-section, wherein the polygonal cross-section is disposed within the fluidic channel;

b) a flow through incubator comprising a first inlet configured to receive separated particles from the first outlet of the microfluidic device, a second inlet fluidically connected to a liquid source comprising a reagent, a serpentine channel, and at least one outlet, and

c) a balancer that is fluidically connected to the second outlet of the microfluidic device such that a liquid exiting from the second outlet of the microfluidic device directly flows into the balancer, wherein the balancer is configured to have a fluidic resistance that is substantially the same as a fluidic resistance through the flow-through incubator,

wherein the balancer comprises a fluidic pressure control comprising a second serpentine channel, and

the fluidic pressure control is capable of generating negative pressure in the balancer.

2 . The system of claim 1 , wherein the serpentine channel of the flow through incubator comprises at least 20 turns.

3 . The system of claim 2 , wherein the reagent is a binding agent that binds to a cell surface marker selected from the group consisting of: CD3; CD4; CD8; CD19; CD20; CD28; CD34; CD38; CD45; and CD56 (wherein “CD” is an abbreviation for cluster of differentiation).

4 . The system of claim 3 , wherein an outlet of said incubator is fluidically connected to an inlet of a second microfluidic device.

5 . The system of claim 1 , wherein said incubator comprises one or more linear channels.

6 . The system of claim 1 wherein the incubator is configured to mix a liquid in the incubator.

7 . The system of claim 1 , wherein the gap defined by the surfaces of said two adjacent obstacles comprises a shape that is substantially symmetrically oriented relative to a plane parallel to the direction of flow of a sample through the array of obstacles and equidistant from the center of the cross-section of each of the two obstacles in the row.

8 . The system of claim 4 , wherein the second microfluidic device comprises a deterministic lateral displacement (DLD) array.

9 . A method for processing first particles of at least a predetermined size from a sample, wherein the first particles comprise leukocytes or stem cells, the method comprising:

a) providing a sample comprising the first particles of at least a predetermined size and second particles of less than the predetermined size; and

b) passing the sample through the system of claim 1 ;

c) separating the first particles from the second particles using the microfluidic device; and

d) incubating the first particles using the flow through incubator by applying the particles separated in b) to the first inlet of the flow through incubator.

10 . The method of claim 9 wherein the reagent flows into said incubator through the second inlet.

11 . The method of claim 10 , wherein the reagent is selected from the group consisting of: a binding agent that binds to a cell surface marker; DNA; an enzyme; a drug; a permeabilization agent and a fixative.

12 . The method of claim 9 , wherein the separated first particles at the first outlet of the microfluidic device comprise at least 80% of the first particles in the sample.

13 . The method of claim 9 , wherein the serpentine channel comprises at least 20 turns.

Assignments (9)

Continuity (2)

References Cited (400)