Self-contained biological assay apparatus, methods, and applications

• US 9,328,381 B2
• Filed: 07/14/2015
• Issued: 05/03/2016
• Est. Priority Date: 10/13/2004
• Status: Active Grant

• Current Assignee: Rheonix, Inc.
• Original Assignee: Rheonix, Inc.
• Inventors: Zhou, Peng, Young, Lincoln C., Thomas, Benjamin, Chen, Zongyuan, Roswech, Todd, Spizz, Gwendolyn, Yasmin, Rubina, Mouchka, Greg
• Primary Examiner(s): Gordon, Brian R
• Application Number:
  US14/798,719
• Publication Number:
  US 20150307921A1
• Time in Patent Office: 294 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: 1/1
• 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 2200/14 Process control and prevention of errors
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0655 pinch valves
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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 3/523 with means for closing or opening
• CPC Class Codes: B01L 3/527 for a plurality of reagents
• 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 1/6809 Methods for determination or identification of nucleic acids involving differential detection
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 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 1/6876 : Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 1/708 : for papilloma
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device
• CPC Class Codes: C12Q 2600/106 : Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
• CPC Class Codes: C12Q 2600/156 : Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/158 : Expression markers
• CPC Class Codes: G01N 2035/00346 : Heating or cooling arrangements
• CPC Class Codes: G01N 35/10 : Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices G01N35/0099 takes precedence
• 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 microfluidic system having a fluidic transport layer and a plurality of reservoirs which are fluidly connected to said fluidic transport layer, wherein the fluidic transport layer, the reservoirs, and a test sample to be introduced into the fluidic transport layer 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,

further wherein the test sample, once introduced, is transported from a starting location in the fluidic transport layer to the analysis reservoir separately from any other samples and separately from the pneumatic manifold and the dispensing system.

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 a 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 4, wherein the reagent storage structure is a length of tubing.

6. The apparatus of claim 2, wherein the reagent dispenser component is a plurality of needle structures of length, L, having different respective, selected bore sizes.

7. The apparatus of claim 6, wherein the reagent dispensing system further includes a plurality of reagent storage structures that are respectively fluidly connected to the plurality of needle structures, wherein each reagent storage structure has a selected dimension that operatively coincides with the respective selected bore size.

8. The apparatus of claim 7, wherein each of the reagent storage structures is a length of tubing.

9. The apparatus of claim 7, having only two reagent storage structures that are respectively fluidly connected to two reagent dispenser components, wherein one of the reagent dispenser components has a bore diameter bi, 0.003<

bi<

0.018 inches and the other reagent dispenser component has a bore diameter b2, 0.015<

b2<

0.030 inches.

10. The apparatus of claim 1, wherein the pneumatic manifold is interfaced with the microfluidic system.

11. The apparatus of claim 10, wherein the microfluidic system further comprises a multi-layer, monolithic, polymeric, non-elastomeric microfluidic component having a given configuration of microfeatures including a plurality of pneumatically-activated diaphragms.

12. The apparatus of claim 11, 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 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 corresponding to a given configuration of the plurality of pneumatically-activated diaphragms in the fluidic transport layer.

13. The apparatus of claim 12, 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 corresponding to the fixed configuration of the plurality of pneumatic-only ports of the pneumatic manifold, removeably connected to the pneumatic manifold.

14. The apparatus of claim 11, wherein the 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 valve disposed in fluid coupling with the at least one reagent reservoir and at least one of the inlet ends,

wherein the 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 valve,

further wherein the multi-layer, monolithic microfluidic component consists of a non-elastomeric, polymeric material.

15. The apparatus of claim 14, wherein the substrate further comprises a plurality of analysis reservoirs, each analysis reservoir including an analysis system disposed therein.

16. The apparatus of claim 14, wherein the analysis system is one of colorimetric, fluorescent colorimetric, chemiluminescent, electrochemical, electrophoretic, lateral flow, protein microarray, nucleic acid microarray, fluorescent.

17. The apparatus of claim 14, further comprising a securing-ring structure having a plurality of perimeter indentations, wherein the analysis membrane is operatively engaged with the securing-ring structure.

18. The apparatus of claim 17, wherein the reservoir has at least a partial inner perimetal shelf upon which the analysis membrane is disposed.

19. The apparatus of claim 17, wherein the securing-ring structure comprises two opposing ring structures each having a plurality of perimeter indentations, further wherein the analysis membrane is disposed intermediate the two opposing ring structures.

20. The apparatus of claim 15, further comprising a heater.

21. The apparatus of claim 20, further comprising a heatable, magnetically engageable reservoir, wherein the heater is disposed adjacent the heatable, magnetically engageable reservoir in such a manner that the heatable, magnetically engageable reservoir can be heated.

22. The apparatus of claim 14, further comprising:

a tube mounting layer attached to a bottom surface of the substrate including a plurality of tubes 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 the tube mounting layer such that the distal end of each tube is disposed substantially near a bottom region thereof.

23. The apparatus of claim 22, further comprising a heater for elevating a temperature in each of the amplification/reaction chambers.

24. The apparatus of claim 11, further comprising a magnetic assembly operably disposed under a reservoir or channel, wherein the magnetic assembly further comprises a magnet, a magnet holder, a piston rod, and a pneumatic piston assembly.

25. The apparatus of claim 24, further comprising a heater assembly operably connected to the magnetic assembly.