Optical lens system and method for microfluidic devices

• US 9,663,821 B2
• Filed: 12/07/2015
• Issued: 05/30/2017
• 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:
  US14/960,754
• Publication Number:
  US 20160194685A1
• Time in Patent Office: 540 Days
• Field of Search: 204602, 204603, 422 681
• 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 nucleic acid sequencing, 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 1,000 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;

maintaining the reaction sites of the fluidic device at a constant temperature;

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

sequencing 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;

reducing chromatic aberration of light along an optical path;

capturing an image of a determined number of reaction sites simultaneously; and

collecting multiple images of the fluidic device, wherein collecting multiple images comprises;

collecting at least a first image at a first wavelength; and

collecting at least a second image at a second wavelength different from the first wavelength.

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 2,500 reaction sites per square centimeter.

3. The method of claim 1, wherein the high-density matrix of reaction sites has a reaction site density of greater than or equal to 4,000 reaction sites per square centimeter.

4. The method of claim 1, wherein each reaction site has a volume of less than 10 nanoliters.

5. The method of claim 1, wherein the electromagnetic radiation emitted by the illumination system has a beam diameter of up to 76 millimeters.

6. The method of claim 1, wherein reducing chromatic aberration is carried out by an optical lens system comprising at least one zero-power optical element coupled to the optical path.

7. The method of claim 1, wherein collecting multiple images further comprises capturing the multiple images over a time frame from about 1 millisecond to about 1 minute.

8. The method of claim 1, wherein collecting multiple images is conducted while maintaining the high-density matrix at a constant temperature.

9. A system for monitoring nucleic acid sequencing, the system comprising:

a fluidic device having a high-density matrix of reaction sites with a reaction site density of greater than or equal to 1,000 reaction sites per square centimeter and configured to receive at least one sample and at least one amplification reagent, wherein the reaction sites of the fluidic device are maintained at a constant temperature during amplification and sequencing of the at least one sample within the high-density matrix;

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

an optical lens system coupled to an optical path and configured to transmit the electromagnetic radiation along the optical path, wherein the optical lens system is configured to reduce chromatic aberration of light along the optical path; and

a detector configured to capture multiple images of the fluidic device, wherein each image simultaneously encompasses a determined number of reaction sites, wherein at least a first image is captured at a first wavelength and at least a second image is captured at a second wavelength different from the first wavelength.

10. The system of claim 9, wherein the high-density matrix of reaction sites has a reaction site density of greater than or equal to 2,500 reaction sites per square centimeter.

11. The system of claim 9, wherein the high-density matrix of reaction sites has a reaction site density of greater than or equal to 4,000 reaction sites per square centimeter.

12. The system of claim 9, wherein each reaction site has a volume of less than 10 nanoliters.

13. The system of claim 9, wherein the illumination system is configured to emit electromagnetic radiation having a beam diameter of up to 76 millimeters.

14. The system of claim 9, wherein the optical lens system comprises at least one zero-power optical element.

15. The system of claim 9, wherein the detector is configured to capture multiple images over a time frame of 1 millisecond to 1 minute.

16. The system of claim 9, wherein the detector is configured to capture multiple images while the high-density matrix is maintained at a constant temperature.