Micro-reactor, biological material inspection device, and microanalysis system

• US 20060094004A1
• Filed: 10/21/2005
• Published: 05/04/2006
• Est. Priority Date: 10/28/2004
• Status: Abandoned Application

• Current Assignee: Konica Minolta Medical & Graphic Incorporated (Konica Minolta Inc.)
• Original Assignee: Konica Minolta Medical & Graphic Incorporated (Konica Minolta Inc.)
• Inventors: Ueda, Eiichi, Higashino, Kusunoki, Ishida, Nobuhisa, Sando, Yasuhiro, Nakajima, Akihisa, Aoki, Youichi
• Application Number:
  US11/255,664
• Publication Number:
  US 20060094004A1
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: B01F 25/4331 Mixers with bended, curved, coiled, wounded mixing tubes or comprising elements for bending the flow
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01L 2200/04 Exchange or ejection of cartridges, containers or reservoirs
• 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 2200/141 Preventing contamination, tampering
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• 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 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 2300/161 Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 2300/1861 using radiation
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0616 Ball valves
• CPC Class Codes: B01L 2400/0638 membrane valves, flap valves
• CPC Class Codes: B01L 2400/0655 pinch valves
• CPC Class Codes: B01L 2400/082 Active control of flow resistance, e.g. flow controllers
• 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 3/502761 : 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 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

Abstract

A micro reactor for inspecting a biological material, comprising: a first substrate on which a minute flow path is formed; a second substrate laminated on the first substrate so as to cover the minute flow path; a detection section provided on the minute flow path formed between the first and second substrates so as to inspect a liquid mixture of a sample and a reagent; (4) an opening section formed at a region on the second substrate corresponding to the detection section so as to irradiate inspection light to the liquid mixture arriving the detection; and (5) a transparent member to cover the opening section.

Claims

1. A micro reactor for inspecting a biological material, comprising:

(1) a first substrate on which a minute flow path is formed;

(2) a second substrate laminated on the first substrate so as to cover the minute flow path;

(3) a detection section provided on the minute flow path formed between the first and second substrates so as to inspect a liquid mixture of a sample and a reagent;

(4) an opening section formed at a region on the second substrate corresponding to the detection section so as to irradiate inspection light to the liquid mixture arriving the detection section; and

(5) a transparent member to cover the opening section.

2. The micro reactor of claim 1, wherein the transparent member is a film, at least the outer periphery of the opening of the second substrate is colored, and the film is heat-sealed on the first substrate by applying heat to this colored area from the film side with a laser.

3. The micro reactor of claim 1, wherein the detection section comprises a land section provided to reduce the depth of the minute flow path.

4. The micro reactor of claim 1, wherein a biotinophilic protein to trap gene amplified in the minute flow path is adsorbed on the detection section.

5. The micro reactor of claim 4, wherein the biotinophilic protein bonds with biotin labeled with 5′

-terminus of a primer used in a gene amplification reaction.

6. The micro reactor of claim 4, wherein the biotinophilic is streptoavidin.

7. The micro reactor of claim 4, wherein the streptoavidin is adsorbed by applying a solution obtained by dissolving the streptoavidin in a buffer solution to the detecting section.

8. The micro reactor of claim 7, wherein a concentration of the streptoavidin in the solution is 10 through 35 μ

g/mL.

9. The micro reactor of claim 7, wherein the buffer solution is a physiological saline solution or an SSC buffer.

10. The micro reactor of claim 4, wherein the amplification reaction is performed according to an ICAN (registered trademark) method.

11. The micro reactor of claim 10, wherein the probe to be hybridized with the amplified gene is labeled with fluorescent pigment.

12. The micro reactor of claim 11, wherein the fluorescent pigment preferably is a FITC.

13. The micro reactor of claim 12, wherein the probe is colored with gold colloid modified by the anti-FITC antibody.

14. The micro reactor of claim 1, wherein biotinophilic protein is absorbed by polystyrene at the detection section to trap a sample gene amplified by a biotin-labeled primer.

15. The micro reactor of claim 14, wherein a probe to be hybridized with the amplified gene is labeled with peroxidase.

16. The micro reactor of claim 15, wherein the probe is made to react with a coloring reagent.

17. The micro reactor of claim 16, wherein the coloring reagent includes as a coloring substance 3,3′

,5,5′

-tetramethyl benzidine, 3,3′

-diamino benzidine, p-phenylene diamine, 5-amino salicylic acid, 3-amino-9-ethylcarbazole, 4-chloro-1-naphthol, 4-amino antipyrine or o-dianisidine.

18. The micro reactor of claim 1, wherein the second substrate to cover the minute flow path and the detection section provide on the first substrate is transparent.

19. The micro reactor of claim 18, wherein a transparent material of the second substrate is polystyrene, a PE, polypropylene, a polyethylene terephthalate, polyethylenenaphthalate, polymethylmethacrylate, polyethylene vinyl alcohol, acryl resin, polyvinyl resin, epoxy resin, polyvinyl chloride, a unsaturated polyester resin, polyamide resin, polyimide resin, polysulfone resin, annular cycloolefin resin, cellulose acetate, cellulose nitrate, fluorocarbon resin, polycarbonate, or poly dimethyl siloxane.

20. The micro reactor of claim 18, wherein a material of the first substrate is polystyrene.

21. The micro reactor of claim 18, wherein an inner surface of the minute flow path is coated with protein.

22. The micro reactor of claim 1, wherein the minute flow path is a minute flow path branched into at least two or more paths, a fluid containing specimen is fed to downstream point in each branched micro flow path and/or a reagent encapsulated or its mixture fluid are fed to downstream points in each branched micro flow path by a micro pump and a feeding fluid dividing means, and a plurality of items and/or controls can be measured simultaneously at downstream points of each branched micro flow path.

23. The micro reactor of claim 22, wherein the micro pump is a piezo pump provided with a first flow path whose flow resistance varies with the change of pressure difference, a second flow path in which a ratio of a change of flow resistance to a change of pressure difference is smaller than that of the first flow path, a pressure chamber connected to both the first flow path and the second flow path, and with an actuator which changes the pressure inside the pressure chamber.

24. The micro reactor of claim 22, wherein the feeding fluid dividing means comprises branched micro flow paths, a feeding fluid control section able to control the passage of the fluid though the pump pressure of the micro pump that intercepts passage of the fluid until the fluid feeding pressure of positive direction reaches to a predetermined pressure and allows passage by applying fluid feeding pressure exceeding the predetermined pressure, and a backward flow preventing section which prevents backward flow in the flow path, and controls feeding of feeding fluid, an fixed amount of feeding fluid and mixing of each fluid in the divided flow path.

25. The micro reactor of claim 1, wherein a substrate is further adhered as a bottom substrate to cover a concave formed on the bottom of the first substrate so as to form a waste fluid reservoir section to reserve waste fluid resulting from condensing, washing and measuring of the specimen.

26. The micro reactor of claim 1, wherein a viscosity of fluid flowing through the minute flow path is controlled to be not more than 10 mPa·

s.