Optical lens system and method for microfluidic devices

• US 10,106,846 B2
• Filed: 04/24/2017
• Issued: 10/23/2018
• 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
• Application Number:
  US15/495,695
• Publication Number:
  US 20170321249A1
• Time in Patent Office: 547 Days
• Field of Search: 422 681, 204602, 204603
• 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 sequencing, the method comprising:

providing a fluidic device comprising;

a plurality of reaction sites having a reaction site density of greater than or equal to 250,000 reaction sites per square centimeter;

at least one sample introduced to the plurality of reaction sites; and

at least one amplification reagent introduced to the plurality of reaction sites;

amplifying the at least one sample at the plurality of reaction sites;

sequencing the at least one amplified sample at the plurality of reaction sites at a predetermined temperature;

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

transmitting the electromagnetic radiation along an optical path coupled to the device and an optical lens system;

reducing chromatic aberration of light with the optical lens system along the 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 plurality of reaction sites has a reaction site density of greater than or equal to 1,000,000 reaction sites per square centimeter.

3. The method of claim 1, wherein reducing chromatic aberration is carried out by at least one zero-power doublet.

4. The method of claim 1, wherein the electromagnetic radiation emitted from the plurality of processing sites is transmitted through a first multi-element optical train and a second multi-element optical train of the optical lens system.

5. 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.

6. The method of claim 1, wherein collecting multiple images is conducted while maintaining the plurality of reaction sites at the predetermined temperature.

7. The method of claim 1, wherein the optical lens system comprises a wheel or filter holder, wherein the filter wheel or holder is positionable to at least two operating positions.

8. The method of claim 1, wherein amplifying the at least one sample comprises:

extending a first polynucleotide sample with a first primer to form a hybrid polynucleotide;

degrading the hybrid polynucleotide to form a second polynucleotide sample; and

extending the second polynucleotide sample with a second primer to form a double-stranded polynucleotide sample.

9. The method of claim 8, wherein amplifying the at least one sample further comprises:

transcribing the double-stranded polynucleotide sample; and

generating multiple anti-sense product of the first polynucleotide sample.

10. The method of claim 1, wherein the electromagnetic radiation transmitted along the optical path comprises electromagnetic radiation emitted from the plurality of processing sites.

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

a fluidic device having a plurality of reaction sites with a reaction site density of greater than or equal to 250,000 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 predetermined temperatures during amplification and sequencing of the at least one sample at the plurality of reaction sites;

a thermal controller coupled to the fluidic device, the thermal controller configured to provide temperature control to the plurality of reaction sites;

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 illumination wavelength and at least a second image is captured at a second illumination wavelength different from the first illumination wavelength.

12. The system of claim 11, wherein the plurality of reaction sites has a reaction site density of greater than or equal to 1,000,000 reaction sites per square centimeter.

13. The system of claim 11, wherein the optical lens system comprises at least one zero-power doublet.

14. The system of claim 11, wherein the detector is configured to collect multiple images over a time frame of 1 millisecond to 1 minute.

15. The system of claim 11, wherein the optical lens system comprises a wheel or filter holder, wherein the filter wheel or holder is positionable to at least two operating positions.

16. The system of claim 11, wherein the optical lens system comprises a first multi-element optical train coupled to a first portion of the optical path and a second multi-element optical train coupled to a second portion of the optical path.

17. The system of claim 11, wherein the fluidic device is configured to amplify nucleic acids to produce an amplified product.

18. The system of claim 17, further comprising sequencing of the amplified product.

19. The system of claim 11, wherein the system is configured to capture the first and second images while maintaining the fluidic device at a predetermined temperature.

20. The system of claim 11, wherein the electromagnetic radiation transmitted along the optical path comprises electromagnetic radiation emitted from the plurality of processing sites.