Split focusing cytometer

• US 6,674,525 B2
• Filed: 04/03/2002
• Issued: 01/06/2004
• Est. Priority Date: 04/03/2001
• Status: Expired due to Term

• Current Assignee: Perkinelmer Health Sciences Incorporated (Perkinelmer Incorporated)
• Original Assignee: Micronics Inc. (Sony Group Corp.)
• Inventors: Weigl, Bernhard H., Battrell, C. Frederick, Bardell, Ronald
• Primary Examiner(s): Font, Frank G.
• Assistant Examiner(s): Nguyen, Sang H.
• Application Number:
  US10/114,790
• Publication Number:
  US 20020149766A1
• Time in Patent Office: 643 Days
• Field of Search: 356/244, 356/246, 356/335-343, 356/39, 356/43, 422/82.05, 422/82.09, 422/100, 422/101, 422/102, 422/55, 422/69, 210/96.1, 210/511, 210/634, 204/452, 204/651
• US Class Current: 356/246
• CPC Class Codes: A61M 1/14 Dialysis systems; Artificial kidneys; Blood oxygenators semi-permeable membranes characterised by the material, manufacturing processes therefor B01D71/00 ; Reciprocating systems for treatment of body fluids, e.g. single needle systems for hemofiltration or pheresis
• CPC Class Codes: A61M 2206/11 Laminar flow
• CPC Class Codes: B01D 21/0012 Settling tanks making use of filters, e.g. by floating layers of particulate material
• CPC Class Codes: B01D 21/283 Settling tanks provided with vibrators
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/028 Modular arrangements
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0436 acoustic forces, e.g. surface acoustic waves [SAW]
• CPC Class Codes: B01L 2400/0457 passive flow or gravitation
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/084 Passive control of flow resistance
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/50273 : characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 : characterised by integrated valves microvalves F16K99/0001
• 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: B01L 3/502776 : specially adapted for focusing or laminating flows
• CPC Class Codes: F16K 2099/008 : Multi-layer fabrications
• CPC Class Codes: F16K 2099/0084 : Chemistry or biology, e.g. "lab-on-a-chip" technology
• CPC Class Codes: F16K 7/17 : the diaphragm being actuated by fluid pressure
• CPC Class Codes: F16K 99/0001 : Microvalves microdevices B81B1/00; manufacture or treatment of devices or systems in or on a substrate B81C1/00; microfluidic structures B01L3/5027; micropumps F04B19/006
• CPC Class Codes: F16K 99/0015 : Diaphragm or membrane valves
• CPC Class Codes: F16K 99/0025 : Valves using microporous membranes
• CPC Class Codes: F16K 99/0059 : actuated by a pilot fluid
• CPC Class Codes: G01N 15/0255 : with mechanical, e.g. inertial, classification, and investigation of sorted collections with centrifuges G01N15/042
• CPC Class Codes: G01N 15/05 : in blood
• CPC Class Codes: G01N 15/1456 : without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
• CPC Class Codes: G01N 2001/4016 : being a selective membrane, e.g. dialysis or osmosis
• CPC Class Codes: G01N 2001/4061 : Solvent extraction
• CPC Class Codes: G01N 2001/4094 : using ultrasound
• CPC Class Codes: G01N 2015/0288 : Sorting the particles
• CPC Class Codes: G01N 2015/1411 : Features of sheath fluids
• CPC Class Codes: G01N 2015/1413 : Hydrodynamic focussing
• CPC Class Codes: G01N 2015/144 : Imaging characterised by its optical setup
• CPC Class Codes: G01N 2015/1486 : Counting the particles
• CPC Class Codes: G01N 2035/00247 : Microvalves
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

A microcytometer which combines lysing and cytometry into a unified system that achieves blood lysis and white blood cell count in a single device. The device focuses the white cells into a thin ribbon which is then focused into a single stream for analysis.

Claims

1. A microfluidic device for analyzing particles dispersed in a sample fluid, comprising:

a first inlet fluidically coupled to a first channel for introduction of the sample fluid;

a second inlet fluidically coupled to a second channel for introduction of a first reagent fluid;

a reactor channel fluidically coupled to the first channel and the second channel such that the sample fluid is focused into a thin ribbon between two streams of the first reagent fluid, the reactor channel having a length sufficient to allow a chemical reaction between the sample fluid and the first reagent fluid;

a third inlet fluidically coupled to a third channel for introduction of a sheath fluid;

a cytometer channel fluidically coupled to the reactor channel and the third channel such that the sample fluid is focused into a single file stream of particles between two streams of the sheath fluid; and

a detection region.

2. The microfluidic device of claim 1, wherein:

the sample fluid comprises whole blood;

the particles are white blood cells; and

the first reagent fluid comprises a lyse reagent.

3. The microfluidic device of claim 2 wherein the lyse reagent is water.

4. The microfluidic device of claim 2 wherein the sheath fluid is phosphate buffered saline.

5. The microfluidic device of claim 1 further comprising a waste chamber coupled to the cytometer channel.

6. A method for analyzing particles dispersed in a sample fluid, comprising:

introducing the sample fluid and a first reagent fluid into a microfluidic device;

focusing the sample fluid into a thin ribbon between two streams of the first reagent fluid;

flowing the sample fluid and first reagent fluid through a reactor channel of the microfluidic device, the reactor channel having a length sufficient to allow a chemical reaction between the sample fluid and the first reagent fluid;

introducing a sheath fluid into the microfluidic device;

focusing the sample fluid into a single file stream of particles between two streams of the sheath fluid; and

flowing the sample fluid, first reagent fluid and sheath fluid through a cytometer channel of the microfluidic device past a detection region of the microfluidic device.

7. The method of claim 6 wherein:

the sample fluid comprises whole blood;

the particles are white blood cells; and

the first reagent fluid comprises a lyse reagent.

8. The method of claim 7 wherein the lyse reagent is water.

9. The method of claim 7 wherein the sheath fluid is phosphate buffered saline.