Mesoscale sample preparation device and systems for determination and processing of analytes

• US 5,726,026 A
• Filed: 11/14/1994
• Issued: 03/10/1998
• Est. Priority Date: 05/01/1992
• Status: Expired due to Term

• Current Assignee: Pennsylvania University of The Trustees of The
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Kricka, Larry J., Wilding, Peter
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): Nguyen, Bao-Thuy L.
• Application Number:
  US08/338,369
• Time in Patent Office: 1,212 Days
• Field of Search: 422/68.1, 422/50, 422/55, 422/58, 435/7.1, 435/91.1, 435/91.2, 435/810, 436/527, 436/538, 436/807
• US Class Current: 435/7.21
• CPC Class Codes: B01D 2325/028 Microfluidic pore structures
• CPC Class Codes: B01D 61/18 Apparatus therefor
• CPC Class Codes: B01D 67/003 by selective elimination of components, e.g. by leaching
• CPC Class Codes: B01D 67/0034 by micromachining techniques, e.g. using masking and etching steps, photolithography
• CPC Class Codes: B01D 67/0058 by selective elimination of components, e.g. by leaching
• CPC Class Codes: B01D 67/0062 by micromachining techniques, e.g. using masking and etching steps, photolithography
• CPC Class Codes: B01D 71/02 Inorganic material
• CPC Class Codes: B01D 71/0215 Silicon carbide; Silicon nitride; Silicon oxycarbide
• CPC Class Codes: B01F 25/41 Mixers of the fractal type
• CPC Class Codes: B01F 35/7182 with means for feeding the material with a fractal or tree-type distribution in a surface
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00828 Silicon wafers or plates
• CPC Class Codes: B01J 2219/00873 Heat exchange
• 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 2300/0681 Filter
• 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 2300/0883 : Serpentine channels
• CPC Class Codes: B01L 2300/1822 : using Peltier elements
• CPC Class Codes: B01L 2300/1827 : using resistive heater
• CPC Class Codes: B01L 2300/1861 : using radiation
• CPC Class Codes: B01L 2400/086 : using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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 7/52 : with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: C12M 41/46 : of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: Y10S 366/03 : Micromixers: variable geometry from the pathway influences mixing/agitation of non-laminar fluid flow
• CPC Class Codes: Y10S 435/81 : Packaged device or kit
• CPC Class Codes: Y10S 436/807 : Apparatus included in process claim, e.g. physical support structures

Abstract

A mesoscale sample preparation device capable of providing microvolume test samples, separated into a cell-enriched fraction and a fraction of reduced cell content, for performing various analyses, such as binding assays, determinations involving polynucleotide amplification and the like. Analytical systems including such devices are also disclosed.

Claims

1. A device for preparing a test sample, comprising particulate components, for analysis, said device comprising a sample flow passage having a sample inlet and an outlet in fluid communication and a separator disposed between said inlet and said outlet, said separator having an upstream-facing portion defining a separation zone in said flow passage in which said particulate components are collected, and a flow channel in fluid communication with said separation zone for affording discharge of collected particulate components from said separation zone, said channel having an inlet section for directing a carrier fluid into said separation zone and over the upstream-facing portion of said separator, and a discharge section for directing said carrier fluid from over the upstream-facing portion of said separator and out of said separation zone, at least one of said flow passage and said flow channel sections having at least one mesoscale dimension.

2. The device of claim 1, wherein said flow passage has at least one mesoscale dimension and said separator comprises a region of restricted flow in said flow passage, said region of restricted flow being formed by at least one passageway having at least one mesoscale dimension smaller than the least mesoscale dimension of said flow passage and being of a small dimension to separate said particulate components from said test sample.

3. The device of claim 2, wherein said at least one passageway has at least one bend therein such that at least a part of said passageway is generally perpendicular to said flow passage.

4. The device of claim 1, wherein said flow passage and said flow channel are formed in a surface of a solid substrate and enclosed by a cover adhered to said surface.

5. The device of claim 4, wherein said separator is in the form of at least one upstanding projection of said solid substrate which is disposed in said flow passage and which restricts the flow of test sample along said flow passage.

6. The device of claim 4, wherein said cover is transparent.

7. In combination, the device of claim 1 and an appliance for use with said device, said appliance comprising a holder for said device, a test sample input conduit interfitted with the sample inlet of said device and an impellent for moving test sample along said flow passage.

8. The combination of claim 7, wherein said appliance further comprises a reservoir for said test sample.

9. The combination of claim 7, wherein said appliance further comprises a carrier fluid input conduit interfitted with said inlet section of said flow channel and an impellent for moving carrier fluid along said flow channel.

10. The combination of claim 9, wherein said appliance further comprises a reservoir for said carrier fluid.

11. A system for determining an analyte in a fluid sample, said system comprising a sample preparation device, according to claim 1, and an analyte detection device comprising:

a solid substrate fabricated to define;

a sample inlet port; and

a flow system comprising;

an analyte detection region in fluid communication with said inlet port, said region containing a reagent which interacts with said analyte to yield a detectable product which is determinative of said analyte, and a detector for detecting said product;

the flow passage outlet of said sample preparation device being in fluid communication with said sample inlet port of said analyte detection device.

12. The system of claim 11, which further comprises, in said analyte detection device, a sample flow channel interconnecting said inlet port and said analyte detection region, at least one of said analyte detection region and said sample flow channel having at least one mesoscale dimension.

13. The system of claim 11, wherein said reagent is a binding substance that binds specifically to said analyte.

14. The system of claim 13, wherein said analyte is an antigen and said binding substance is an antibody.

15. The system of claim 13, wherein said analyte is a ligand and said binding substance is a receptor.

16. The system of claim 13, wherein said analyte is a nucleic acid molecule of predetermined sequence and said binding substance is a nucleic acid molecule having a sequence complementary or homologous to the sequence of said analyte.

17. The system of claim 11 further comprising a device for performing analysis of preselected polynucleotide derived from cells, said analysis comprising a polynucleotide amplification reaction, said polynucleotide analysis device comprising:

a solid substrate fabricated to define;

a sample inlet port; and

a flow system comprising;

a polynucleotide amplification region in fluid communication with said inlet port, said polynucleotide amplification region containing reagents for amplifying a polynucleotide, and lysing means intermediate the discharge section of the flow channel of said sample preparation device and said polynucleotide amplification region for lysing said cells, the discharge section of the flow channel of said sample preparation device being in fluid communication with the sample inlet port of said polynucleotide analysis device.

18. The system of claim 17, which further comprises a sample flow channel in said polynucleotide analysis device interconnecting the inlet port of said polynucleotide amplification device and said polynucleotide amplification region at least one of said polynucleotide amplification region and the sample flow channel connected therewith having at least one mesoscale dimension.

19. In combination, the system of claim 11 and an appliance for use with said system, said appliance comprising a holder for said system, a test sample input conduit interfitted with the sample inlet of said sample preparation device and an impellent for moving test sample along the flow passage of said sample preparation device.

20. The combination of claim 19, wherein said appliance further comprising a reservoir for said test sample.

21. In combination, the system of claim 17, and an appliance for use with said system, said appliance comprising a holder for said system, a test sample input conduit interfitted with the sample inlet of said sample preparation device, and an impellent for moving said test sample along the flow passage of said sample preparation device, a carrier fluid input conduit interfitted with said inlet section of the flow channel of said sample preparation device and an impellent for moving said carrier fluid along said flow channel.

22. The combination of claim 21, wherein said appliance further comprises a reservoir for the carrier fluid which is directed into the separation zone of said sample preparation device.

23. The combination of claim 21, wherein said appliance further comprises a detector for detecting a parameter of said test sample in said analyte detection device or said polynucleotide analysis device.

24. A system for performing an analysis of preselected polynucleotide derived from cells, said analysis comprising polynucleotide amplification, said system comprising a sample preparation device according to claim 1 and a device for carrying out polynucleotide amplification which comprises:

a solid substrate fabricated to define;

a sample inlet port; and

a flow system comprising;

a polynucleotide amplification region in fluid communication with said inlet port, said polynucleotide amplification region containing reagents for amplifying a polynucleotide, and a sample flow channel in said polynucleotide amplification device interconnecting the inlet port of said polynucleotide amplification device and said polynucleotide amplification region, at least one of said sample flow channel and said polynucleotide amplification region having at least one mesoscale dimension, and lysing means in said flow channel upstream of said polynucleotide amplification region for lysing said cells, the discharge section of the flow channel of said sample preparation device being in fluid communication with the sample inlet port of said polynucleotide amplification device.

25. The system of claim 24, which further comprises, in said polynucleotide amplification device, a sample flow channel of said polynucleotide amplification device interconnecting said inlet port and said polynucleotide amplification region, at least one of said polynucleotide amplification region and the sample flow channel connected thereto having at least one mesoscale dimension.

26. A system for determining an analyte in a fluid sample having particulate components, said system comprising a sample preparation device which comprises a sample flow passage having a sample inlet and an outlet in fluid communication and a separator disposed between said inlet and said outlet, said separator having an upstream-facing portion defining a separation zone in said flow passage in which said particulate components are collected, and an analyte detection device comprising:

a solid substrate fabricated to define;

a sample inlet port; and

a flow system comprising;

an analyte detection region in fluid communication with said inlet port, said region containing a reagent which interacts with said analyte to yield a detectable product which is determinative of said analyte, and a detector for detecting said product;

the flow passage outlet of said sample preparation device being in fluid communication with said sample inlet port of said analyte detection device, and at least one of said flow passage and said region having at least one mesoscale dimension.

27. The system of claim 26, which further comprises, in said sample preparation device, a flow channel in fluid communication with said separation zone for affording discharge of collected particulate components from said separation zone, said flow channel having an inlet section for directing a carrier fluid into said separation zone and over the upstream-facing portion of said separator, and a discharge section for directing said carrier fluid from over the upstream-facing portion of said separator and out of said separation zone.

28. The system of claim 27, wherein at least one of the inlet and discharge sections of said flow channel has at least one mesoscale dimension.