Thermo-controllable chips for multiplex analyses

• US 7,833,709 B2
• Filed: 05/24/2005
• Issued: 11/16/2010
• Est. Priority Date: 05/28/2004
• Status: Active Grant

• Current Assignee: Takara Bio Usa, Inc. (Ningbo Junsheng Investment Group Co., Ltd.)
• Original Assignee: WaferGen, Inc.
• Inventors: Zhou, Jie, Joseph, Victor, Huda, Amjad, Shivji, Alnoor
• Primary Examiner(s): Beisner; William H
• Assistant Examiner(s): HOBBS, MICHAEL L
• Application Number:
  US11/136,756
• Publication Number:
  US 20060030035A1
• Time in Patent Office: 2,002 Days
• Field of Search: 435/6, 435/288.4, 435/288.7, 435/303.1
• US Class Current: 435/6.14
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 3/50853 with covers or lids
• 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 7/54 using spatial temperature gradients
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• 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 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2565/501 being an array of oligonucleotides
• CPC Class Codes: G01N 2021/6417 Spectrofluorimetric devices
• CPC Class Codes: G01N 2021/6484 Optical fibres
• CPC Class Codes: G01N 2035/0405 : manipulating closing or opening means, e.g. stoppers, screw caps, lids or covers
• CPC Class Codes: G01N 21/01 : Arrangements or apparatus for facilitating the optical investigation
• CPC Class Codes: G01N 21/0332 : with temperature control control of temperature G05D23/00; cryostats F17C3/08
• CPC Class Codes: G01N 21/6408 : with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 21/6452 : Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/76 : Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 2201/0231 : Thermostating
• CPC Class Codes: G01N 35/0099 : comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: H01L 27/1443 : with at least one potential jump or surface barrier
• CPC Class Codes: H01L 31/0203 : Containers; Encapsulations , e.g. encapsulation of photodiodesfor photovoltaic devices H01L31/048; for organic photosensitive devices H10K30/80
• CPC Class Codes: H01L 31/02325 : the optical elements not being integrated nor being directly associated with the device
• CPC Class Codes: H01L 31/024 : Arrangements for cooling, heating, ventilating or temperature compensation for photovoltaic devices H01L31/052
• CPC Class Codes: H01L 31/105 : the potential barrier being of the PIN type
• CPC Class Codes: H01L 31/18 : Processes or apparatus specially adapted for the manufacture or treatment of these devices or of parts thereof

Abstract

The present invention provides miniaturized instruments for conducting chemical reactions where control of the reaction temperature is desired or required. Specifically, this invention provides chips and optical systems for performing and monitoring temperature-dependent chemical reactions. The apparatus and methods embodied in the present invention are particularly useful for high-throughput and low-cost amplification of nucleic acids.

Claims

1. A method of performing a nucleic acid amplification reaction using a miniaturized nucleic acid amplification chip, said amplification involving a plurality of reaction samples and requiring cycling at least two temperature levels, comprising:

(a) providing a chip comprising an array of fluidically isolated micro wells, each of said micro wells is configured for receiving and confining a reaction sample, wherein the micro well is coated with a hydrophobic coating, and wherein the chip is placed in thermal contact with a heating element;

(b) dispensing the plurality of reaction samples into the fluidically isolated micro wells in ambient air that is saturated with moisture with respect to a planar surface of the chip using a dispenser under conditions to reduce evaporation of said reaction samples;

(c) controlling the heating element to effect cycling of the at least two temperature levels; and

(d) detecting optical signals coming from the reaction samples using an optical detection assembly, wherein the optical signals are indicative of the presence of amplified nucleic acids.

2. The method of claim 1, wherein the controlling step comprises processing sensor signals retrieved from a temperature sensor operatively linked to a micro well based on a protocol stored on a computer readable medium.

3. The method of claim 1, wherein the optical signals are proportional to the amount of the amplified nucleic acids.

4. The method of claim 1, wherein the chip further comprises individually-controlled heating elements, each of which is operably coupled to a micro well of said array.

5. The method of claim 1 further comprising sealing a micro well by applying an adhesive layer over an open surface of the micro well.

6. The method of claim 5 further comprising applying a cover over the adhesive layer wherein the cover is transparent and comprises an ITO heater.

7. The method of claim 6 wherein the ITO heater heats the cover to a temperature greater than at least one of the two temperature levels.

8. The method of claim 1 wherein the chip has a ramp temperature time of about 25 degrees C. or higher.

9. The method of claim 1 wherein the plurality of reaction samples are dispensed into the micro wells while said micro wells are kept at about dew point temperature.

10. The method of claim 1 wherein the plurality of reaction samples are dispensed into the micro wells while said dispensing is carried out at temperatures no more than 1-5 degrees C. over dew point.

11. The method of claim 1 wherein the optical signal is a fluorescent signal.

12. The method of claim 1 wherein reaction samples comprise reagents necessary for said nucleic acid amplification reaction and a detectable label, wherein said detectable label generates said optical signal that is detected.

13. The method of claim 1 wherein said nucleic acid amplification reaction is polymerase chain reaction (PCR).

14. The method of claim 1 wherein the optical signals coming from the reaction samples are monitored during said nucleic acid amplification reaction.

15. The method of claim 1 wherein said nucleic acid amplification reaction is real-time polymerase chain reaction.

16. The method of claim 1 wherein the micro well has a volume of approximately 0.001 μ

l to 100 μ

l.

17. The method of claim 1 wherein the micro well has a volume of approximately 1 μ

l to 10 μ

l.

18. The method of claim 1 wherein the micro well has a dimension of approximately 100 μ

m to 10 mm in length by 100 μ

M to 10 mm in width.