Chemiluminescence-based microfluidic biochip

• US 6,949,377 B2
• Filed: 12/13/2001
• Issued: 09/27/2005
• Est. Priority Date: 03/05/2001
• Status: Expired due to Fees

• Current Assignee: Winston Z. Ho
• Original Assignee: Winston Z. Ho
• Inventors: Ho, Winston Z.
• Primary Examiner(s): Horlick, Kenneth R.
• Assistant Examiner(s): LU, FRANK WEI MIN
• Application Number:
  US10/022,007
• Publication Number:
  US 20020123059A1
• Time in Patent Office: 1,384 Days
• Field of Search: 435/6.1, 435/91.1, 435/183, 435/283.1, 435/287.1, 435/287.2, 435/287.3, 435/288.4, 435/288.5, 435/292.1, 422/50, 422/68.1, 422/82.05, 422/82.07, 422/201, 530/300, 530/350, 536/23.1, 424/130.1, 424/184.1
• US Class Current: 435/287.1
• 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 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• 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 2300/0864 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 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0683 mechanically breaking a wall or membrane within a channel or chamber
• 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 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 21/7703 using reagent-clad optical fibres or optical waveguides using measurement of total internal reflection or attenuated total reflection G01N21/552; optical fibres or waveguides per se G02B

Abstract

The disclosure describes how to use luminescence detection mechanism, move microfluid, and control multiple-step biochemical reactions in closed confined microfluidic biochip platform. More particularly, a self-contained disposable biochip with patterned microchannels and compartments having storage means for storing a plurality of samples, reagents, and luminescent substrates. At least one external microactuator in the biochip system produces positive pressure and automates multiple-step reactions in microfluidic platforms for clinical chemistry, cell biology, immunoassay and nucleic acid analysis. The method comprises the steps of transferring sequentially at least one of samples, reagents, and then luminescent substrate from compartments through microchannels to reaction sites. The luminescent substrates react with probes to form a probe complex resulting into luminescence, which is detected by an optical detector.

Claims

1. A method for performing biological assay in a self-contained microfluidic-based luminescence biochip, the method comprising the steps of:

(a) providing said biochip with a plurality of compartments and interconnecting microchannels, said compartments having storage means for storing a plurality of samples, reagents, luminescent substrates, and reaction sites immobilized with probes; and

means for interconnecting said compartments to provide fluid transfer through said microchannels;

(b) transferring sequentially at least one of said samples and then at least one of said reagents through said microchannels to at least one of said reaction sites, said at least one sample and said at least one reagent reacting with said probes and forming a probe complex;

(c) transferring said luminescent substrate through said microchannels to said reaction sites, said luminescent substrates reacting with said probe complex and generating luminescence; and

(d) detecting said luminescence with an optical detector located above or under said reaction sites.

2. The method as defined in claim 1, wherein said compartments further include washing buffers, the method further including a step prior to said step (c), said step comprising transferring said washing buffers through said microchannels to said reaction sites and washing away an excessive and un-reacting portion of said samples or said reagents.

3. The method as defined in claim 1 or 2, wherein one of said luminescent substrates is a chemiluminescent material or a bioluminescence material.

4. The method as defined in claim 1 or 2, wherein one of said reagents is an enzyme conjugate.

5. The method as defined in claim 1 or 2, wherein at least one of said samples is selected from a group consisting of proteins, antibodies, antigens, hormones, biological cells, and oligonucleotides.

6. The method as defined in claim 1 or 2, wherein one of said reagents is a branched DNA amplifier adapted for luminescence signal amplification.

7. The method as defined in claim 1 or 2, wherein means for interconnecting said compartments to provide fluid transfer is based on an external microactuator positioned above said compartments.

8. The method as defined in claim 1 or 2, wherein said microchannels have a dimension between 10 μ

m-3 mm.

9. The method as defined in claim 1 or 2, wherein said compartments have a volume between 100 nl-500 μ

l.

10. The method as defined in claim 1 or 2, wherein said compartments formed by thin film materials.

11. The method as defined in claim 1 or 2, wherein one of said reagents is dry reagent.