Optical lens system and method for microfluidic devices

• US 9,234,237 B2
• Filed: 11/19/2014
• Issued: 01/12/2016
• Est. Priority Date: 06/07/2004
• Status: Active Grant

• Current Assignee: Standard Biotools Incorporated
• Original Assignee: Fluidigm Corporation
• Inventors: Unger, Marc A., Facer, Geoffrey Richard, Clerkson, Barry, Cesar, Christopher G., Switz, Neil
• Primary Examiner(s): CHOI, WILLIAM C
• Application Number:
  US14/547,442
• Publication Number:
  US 20150203896A1
• Time in Patent Office: 419 Days
• Field of Search: 204/602, 204/603, 422/68.1
• US Class Current: 1/1
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00704 integrated with the reactor apparatus
• 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 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 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 21/75 Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated systems in which material is burnt in a flame or plasma G01N21/72, G01N21/73
• CPC Class Codes: G01N 21/8483 Investigating reagent band test-element handling not specific to a test method G01N33/4875; analytical elements specific to chemical analysis of biological material G01N33/52; autometer with reagent band G01N35/04
• CPC Class Codes: G01N 2201/061 Sources
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G02B 21/16 adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence
• CPC Class Codes: G02B 21/36 arranged for photographic purposes or projection purposes G02B21/18 takes precedenceor digital imaging or video purposes including associated control and data processing arrangements image data processing per se G06T

Abstract

An apparatus for imaging one or more selected fluorescence indications from a microfluidic device. The apparatus includes an imaging path coupled to least one chamber in at least one microfluidic device. The imaging path provides for transmission of one or more fluorescent emission signals derived from one or more samples in the at least one chamber of the at least one microfluidic device. The chamber has a chamber size, the chamber size being characterized by an actual spatial dimension normal to the imaging path. The apparatus also includes an optical lens system coupled to the imaging path. The optical lens system is adapted to transmit the one or more fluorescent signals associated with the chamber.

Claims

1. A method of monitoring DNA amplification, the method comprising:

providing a fluidic device comprising;

a high-density matrix of reaction sites having a reaction site density of greater than or equal to 10,000 reaction sites per square centimeter;

at least one sample introduced into the high-density matrix; and

at least one amplification reagent introduced into the high-density matrix;

thermally controlling at a constant temperature the high density of reaction sites of the fluidic device with a thermal controller;

amplifying the at least one sample within the high-density matrix at the constant temperature;

illuminating the fluidic device with electromagnetic radiation with an illumination system; and

collecting multiple images of the fluidic device, wherein each image collected comprises an image of a determined number of the reaction sites, and wherein collecting multiple images comprises;

reducing chromatic aberration of multiple images along an optical path;

collecting at least a first image at a first wavelength;

collecting at least a second image at a second wavelength different from the first wavelength; and

collecting each image of the determined number of the reaction sites simultaneously while maintaining the constant temperature of the determined number of reaction sites with the thermal controller.

2. The method of claim 1, wherein the high-density matrix of reaction sites has a reaction site density of greater than or equal to 25,000 reaction sites per square centimeter.

3. The method of claim 1, wherein the reaction site density is greater than or equal to 100,000 reaction sites per square centimeter.

4. The method of claim 1, wherein the reaction site density is greater than or equal to 250,000 reaction sites per square centimeter.

5. The method of claim 1, wherein the reaction site density is greater than or equal to 1,000,000 reaction sites per square centimeter.

6. The method of claim 1, wherein illuminating the fluidic device with electromagnetic radiation further comprises providing multiple wavelengths of electromagnetic radiation.

7. The method of claim 6, wherein the electromagnetic radiation illuminating the fluidic device has an optical path that passes through one or more spectral filters.

8. The method of claim 6, wherein the one or more spectral filters include a filter wheel along the optical path.

9. The method of claim 1, wherein amplifying the at least one sample within the high-density matrix at the constant temperature further comprises:

performing nucleic acid amplification in at least the determined number of reaction sites in the high-density matrix; and

regulating the nucleic acid amplification with the thermal controller.

10. The method of claim 1, further comprising quantifying contents of the determined number of reaction sites of the fluidic device using the multiple images.

11. A system for monitoring DNA amplification, the system comprising:

a fluidic device including a high-density matrix of reaction sites having a reaction site density of greater than or equal to 10,000 reaction sites per square centimeter, configured to receive at least one sample and at least one amplification reagent, and configured to accommodate amplification of the at least one sample at the reaction sites;

a thermal controller configured to maintain the high density of reaction sites of the fluidic device at a constant temperature;

an illumination system configured to illuminate the fluidic device with electromagnetic radiation; and

a detector configured to collect multiple images of the fluidic device, the detector configured to reduce chromatic aberration of the multiple images along an optical path, wherein each image collected comprises an image of a determined number of the reaction sites, wherein each of the multiple images collected comprise at least a first image collected at a first illumination wavelength and at least a second image collected at a second illumination wavelength different from the first illumination wavelength, and wherein each image is collected simultaneously while maintaining the constant temperature of the determined number of reaction sites with the thermal controller.

12. The system of claim 11, wherein the reaction site density is greater than or equal to 25,000 reaction sites per square centimeter.

13. The system of claim 11, wherein the reaction site density is greater than or equal to 100,000 reaction sites per square centimeter.

14. The system of claim 11, wherein the reaction site density is greater than or equal to 250,000 reaction sites per square centimeter.

15. The system of claim 11, wherein the reaction site density is greater than or equal to 1,000,000 reaction sites per square centimeter.

16. The system of claim 11, wherein the electromagnetic radiation has an optical path that passes through at least one filter configured to filter at least one illumination wavelength of the electromagnetic radiation.

17. The system of claim 16, further comprising an excitation filter positioned between the illumination system and the fluidic device along the optical path of the electromagnetic radiation.

18. The system of claim 16, further comprising an emission filter positioned between the fluidic device and the detector along the optical path of the electromagnetic radiation.

19. The system of claim 16, wherein the at least one filter further comprises a filter wheel.

20. The system of claim 11, wherein the thermal controller is configured to maintain a constant temperature during amplification of the at least one sample located within the determined number of the reaction sites.