Self-contained biological assay apparatus, methods, and applications

• US 8,383,039 B2
• Filed: 02/23/2011
• Issued: 02/26/2013
• Est. Priority Date: 02/23/2010
• Status: Active Grant

• Current Assignee: Rheonix, Inc.
• Original Assignee: Rheonix, Inc.
• Inventors: Zhou, Peng, Young, Lincoln C., Thomas, Benjamin, Chen, Zongyuan, Mouchka, Greg, Roswech, Todd, Spizz, Gwendolyn, Yasmin, Rubina
• Primary Examiner(s): Gordon, Brian R
• Application Number:
  US13/033,165
• Publication Number:
  US 20110275058A1
• Time in Patent Office: 734 Days
• Field of Search: 422500-505, 422507-509, 422/518, 422 62- 67, 422/68.1, 422/400, 422 8205- 8209, 436/43, 436/49, 436/164, 436/166, 436/180
• US Class Current: 422/63
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/141 Preventing contamination, tampering
• CPC Class Codes: B01L 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0838 Capillaries
• 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 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/088 Channel loops
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0666 Solenoid valves
• CPC Class Codes: B01L 3/022 Capillary pipettes, i.e. having very small bore B01L3/0224 - B01L3/0237 take precedence
• 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 3/502753 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: C12Q 1/6806 : Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6846 : Common amplification features
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6883 : for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 1/708 : for papilloma
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device
• CPC Class Codes: G01N 35/1002 : Reagent dispensers

Abstract

A self-contained, fully automated, biological assay-performing apparatus includes a housing; a dispensing platform including a controllably-movable reagent dispensing system, disposed in the housing; a reagent supply component disposed in the housing; a pneumatic manifold removably disposed in the housing in a space shared by the dispensing platform, removably coupled to a fluidic transport layer and a plurality of reservoirs, wherein the fluidic transport layer, the reservoirs, and a test sample to be introduced therein are disposed in the housing in the space separate from the dispensing platform; a pneumatic supply system removably coupled to the pneumatic manifold in the housing in a space separate from the dispensing platform; and a control system coupled to at least one of the dispensing platform and the pneumatic supply system, disposed in the housing.

Claims

1. A self-contained, biological assay apparatus, comprising:

a housing;

a dispensing platform including a controllably-movable reagent dispensing system, disposed in the housing;

a reagent supply component disposed in the housing;

a pneumatic manifold removeably disposed in the housing in a space shared by the dispensing platform, removeably coupled to a fluidic transport layer and a plurality of reservoirs,

wherein the fluidic transport layer, the reservoirs, and a test sample to be introduced therein are disposed in the housing in the space separate from the dispensing platform;

a pneumatic supply system removeably coupled to the pneumatic manifold in the housing in a space separate from the dispensing platform; and

a control system coupled to at least one of the dispensing platform and the pneumatic supply system, disposed in the housing,

wherein the apparatus is characterized as a self-contained, fully automated biological assay-performing apparatus.

2. The apparatus of claim 1, wherein the dispensing platform further comprises:

a motion control system operatively coupled to the reagent dispensing system,

wherein the reagent dispensing system includes a reagent dispenser component having a distal dispensing end; and

a camera connected to the reagent dispensing system having a field of view that includes at least a selected region of interest of the reservoirs.

3. The apparatus of claim 2, wherein the reagent dispenser component is an elongate needle structure having a length, L, and a selected bore size.

4. The apparatus of claim 3, wherein the reagent dispensing system further includes a reagent storage structure fluidly connected to the reagent dispenser component, wherein the reagent storage structure has a selected dimension that operatively coincides with the selected bore size.

5. The apparatus of claim 2, wherein the reagent dispenser component is a pipette tip.

6. The apparatus of claim 1, wherein the pneumatic manifold is interfaced with a microfluidic system having at least one assay capacity and wherein the microfluidic system further comprises a two-layer monolithic, polymeric, non-elastomeric microfluidic component having a given configuration of microfeatures including a plurality of pneumatically-activated diaphragms.

7. The apparatus of claim 6, further comprising an operable magnetic assembly disposed under a reservoir or channel.

8. The apparatus of claim 6, wherein the pneumatic manifold has a plurality of pneumatic only ports on an underside thereof, and a plurality of pneumatic only channels disposed therein in fluid connection with a plurality of diaphragm valves in the fluidic transport layer and the plurality of pneumatic only ports, wherein said plurality of pneumatic ports have a fixed configuration, and said plurality of pneumatic only channels have a given configuration specifically corresponding to a given configuration of the plurality of pneumatically-activated diaphragms in the fluidic transport layer.

9. The apparatus of claim 8, wherein the pneumatic supply system further comprises a plurality of aperture tubes that provide a passage of the pneumatic signal there through, in fluid connection with said plurality of pneumatic only ports, wherein said plurality of aperture tubes have a fixed configuration specifically corresponding to the fixed configuration of the plurality of pneumatic only ports of the pneumatic manifold, removeably connected to the pneumatic manifold.

10. The apparatus of claim 6, wherein each multi-layer, monolithic microfluidic component further comprises:

a polymeric, non-elastomeric substrate having a plurality of fluid channels disposed therein, each of the fluid channels having an inlet end and an outlet end;

at least one reagent reservoir of a type capable of holding a reagent material;

at least one bi-directional diaphragm pump comprising at least three non-elastomeric membrane-based diaphragm valve structures; and

a diaphragm valve disposed in fluid coupling with the at least one reagent reservoir and at least one of the inlet ends,

wherein the diaphragm valve is adapted to controllably direct a flow of the material from the at least one reagent reservoir to a plurality of reservoirs via at least one of the channels coupled to the diaphragm valve,

further wherein each two-layer, monolithic microfluidic component consists of a nonelastomeric, polymeric material.

11. The apparatus of claim 10, wherein each substrate further comprises at least one analysis reservoir, each analysis reservoir including an analysis system disposed therein wherein the analysis system is one of colorimetric, fluorescent colorimetric, chemiluminescent, electrochemical, electrophoretic, lateral flow, protein microarray, nucleic acid microarray, fluorescent.

12. The apparatus of claim 10, further comprising:

a plurality of lumens attached to a bottom surface of the substrate each having a proximal end that is fluidly connected to a respective fluidic channel and a distal end projecting downwardly perpendicularly from the substrate; and

a respective plurality of amplification/reaction chambers attached at a top region thereof to a film layer such that the distal end of each lumen is disposed within the amplification/reaction chamber and disposed substantially near a bottom region thereof.

13. The apparatus of claim 12, further comprising at least one heater incorporated into the pneumatic manifold and interfitting with the exterior of the distal end of the amplification/reaction chamber for elevating a temperature in each of the amplification/reaction chambers.