Method to analyze a sample fluid in a microfluidic cytometry system

• US 8,735,088 B2
• Filed: 07/06/2010
• Issued: 05/27/2014
• Est. Priority Date: 07/07/2009
• Status: Active Grant

• Current Assignee: Sony Corporation (Sony Group Corp.), Sony Corporation Of America (Sony Group Corp.)
• Original Assignee: Sony Corporation (Sony Group Corp.), Sony Corporation Of America (Sony Group Corp.)
• Inventors: Durack, Gary P.
• Primary Examiner(s): Weber, Jon P
• Assistant Examiner(s): Srivastava, Kailash C
• Application Number:
  US12/830,876
• Publication Number:
  US 20110008818A1
• Time in Patent Office: 1,421 Days
• Field of Search: 422/72, 435/29
• US Class Current: 435/29
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• 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 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• 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 1/4077 by other techniques involving separation of suspended solids
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence

Abstract

The present disclosure relates to microfluidic devices adapted for post-centrifugation use with selective sample extraction, and methods for their use. Certain embodiments make use of a dye-selective sample extraction. Other embodiments make use of a geographically-selective sample extraction. Other embodiments are also disclosed.

Claims

1. A method of analyzing a sample fluid in a microfluidic cytometry system, the method comprising the steps of:

a) supplying a sample to a sample well in a microfluidic device;

b) centrifuging the microfluidic device to create a first sample layer and a second sample layer within said sample well;

c) extracting first fluid from said first sample layer; and

d) analyzing said extracted first fluid using cytometry while said extracted first fluid is in the microfluidic device.

2. The method of claim 1, further comprising the steps of:

e) sorting a first portion of said extracted first fluid into a first well based upon results obtained in step (d); and

f) sorting a second portion of said extracted first fluid into a second well based upon results obtained in step (d).

3. The method of claim 2, wherein said sorting at steps (e) and (f) is selected from the group consisting of:

greater than a binary sort, greater than a tertiary sort, greater than a quaternary sort.

4. The method of claim 2, wherein said sorting at steps (e) and (f) employs a predetermined number of microfluidic channels, said predetermined number selected from the group consisting of:

at least five channels, at least ten channels, at least 100 channels, and at least 256 channels.

5. The method of claim 2, further comprising the steps of:

g) extracting second fluid from said second sample layer; and

h) analyzing said extracted second fluid using cytometry while said extracted second fluid is in the microfluidic device.

6. The method of claim 5, further comprising the step of:

i) sorting a third portion of said extracted second fluid into a third well based upon results obtained in step (h).

7. The method of claim 5, wherein:

said extracted first fluid is extracted from said sample well at a first location; and

said extracted second fluid is extracted from said sample well at a second location, said second location being different than said first location.

8. The method of claim 1, further comprising the step of:

adding a chemical additive to said sample well prior to performing step (a).

9. The method of claim 1, further comprising the step of:

adding Ficoll-Hypaque to said sample well prior to performing step (a).

10. The method of claim 1, further comprising the step of:

adding a dye to said sample well prior to performing step (a).

11. The method of claim 1, wherein step (a) further comprises supplying a sample of human blood to a sample well on a microfluidic device.