Device and method for extracting individual picoliter droplets from microfluidic emulsions for further analysis and scale-up
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
B01L-003/00
G01N-035/10
B01L-003/02
G01N-015/14
C12M-001/00
C12M-001/12
출원번호
US-0937672
(2015-11-10)
등록번호
US-10040067
(2018-08-07)
우선권정보
DE-10 2014 116 357 (2014-11-10)
발명자
/ 주소
Tovar, Miguel
Zang, Emerson
Weber, Thomas
출원인 / 주소
LEIBNIZ-INSTITUT FÜR NATURSTOFF-FORSCHUNG UND INFEKTIONSBIOLOGIE—HANS-KNÖLL-INSTITUT
대리인 / 주소
Knobbe, Martens, Olson & Bear, LLP
인용정보
피인용 횟수 :
0인용 특허 :
4
초록▼
The disclosure relates to a method and a system for the isolation of picoliter droplets from a continuous stream of carrier fluid in a microfluidic system. In particular the disclosure relates to a system that comprises a sorter, a detector and a collector. The sorter serves for a selection of picol
The disclosure relates to a method and a system for the isolation of picoliter droplets from a continuous stream of carrier fluid in a microfluidic system. In particular the disclosure relates to a system that comprises a sorter, a detector and a collector. The sorter serves for a selection of picoliter droplets with a desired content based upon an optical signal obtained from the picoliter droplets. The selected picoliter droplets are guided towards the detector that comprises a detection channel that is arranged perpendicular to a light beam. By means of a photodetector that is aligned with the optical axis of the light beam the passage of the picoliter droplets can be detected. The collector comprises multiple wells and the detection of the passage of the picoliter droplets results in a relative movement of the detector and a collector such that the picoliter droplet is isolated by depositing it into one of said wells. The disclosure moreover relates to a method that uses such a system to isolate individual picoliter droplets. In particular the disclosure relates to a method that allows for an analysis and/or further processing of the content of the picoliter droplets that have been isolated from a microfluidic system by depositing them into a well.
대표청구항▼
1. A system for isolating picoliter droplets comprising: a) a sorter comprising a microfluidic chip configured for the passage of picoliter droplets and their selection based upon an optical signal obtained from the picoliter droplets, wherein the sorter comprises a first light source and a camera o
1. A system for isolating picoliter droplets comprising: a) a sorter comprising a microfluidic chip configured for the passage of picoliter droplets and their selection based upon an optical signal obtained from the picoliter droplets, wherein the sorter comprises a first light source and a camera or a first photodetector such that the picoliter droplets are sorted based upon the optical signal obtained by the camera or the first photodetector,b) a detector comprising a detection channel with an inner cross-sectional dimension smaller than 150 μm, a second light source generating a light beam and a second photodetector, wherein the detection channel is arranged perpendicular to the optical axis of the light beam in between the second light source and the second photodetector, such that the passage of a picoliter droplet is detected by a change of the refraction angle of said light beam upon which the second photodetector outputs a signal, andc) a collector comprising multiple wells,wherein the detector and the collector are configured to move in relation to each other based upon the signal generated at the second photodetector, and wherein detection of the passage of the picoliter droplet results in a relative movement of the detector and the collector, such that the picoliter droplet is isolated by depositing it into one of the multiple wells. 2. The system according to claim 1, wherein the microfluidic chip comprises microfluidic channels that form a Y-shaped junction and/or a pair of electrodes configured for a dielectrophoretic sorting of the droplets. 3. The system according to claim 1, wherein the picoliter droplets are sorted based upon an analysis of an image of a picoliter droplet detected by the camera. 4. The system according to claim 1, wherein the picoliter droplets, whereby the picoliter droplets are sorted based upon a fluorescent signal of a picoliter droplet detected by the first photodetector. 5. The system according to claim 1, wherein the detection channel of the detector is a capillary. 6. The system according to claim 1, wherein the system comprises a gas inlet into the detection channel, which is situated below the optical axis of the light beam. 7. The system according to claim 5, wherein the capillary is fixed within a capillary guider comprising two connectors arranged on the optical axis of the light beam, wherein the first connector is configured to receive a first optical fiber that extends from the second light source and the second connector is configured to receive a second optical fiber that extends towards the second photodetector. 8. A method for isolating picoliter droplets comprising: 1) providing a system according to claim 1,2) transporting a picoliter droplet in a carrier fluid towards the sorter,3) selecting a picoliter droplet within the microfluidic chip based upon an optical signal obtained from said picoliter droplet,4) transporting the selected picoliter droplet towards the detection channel,5) detecting the passage of a selected picoliter droplet through the light beam in the detection channel due to a change of light intensity measured at the first photodetector,6) triggering a movement of the detection channel and/or the collector when a passage of a picoliter droplet has been detected in step 5), and7) depositing the picoliter droplet into a well. 9. The method according to claim 8, wherein the picoliter droplet comprises an aqueous buffer and a biological material selected from a group consisting of eukaryotic cells, prokaryotic cells, plant cells, viruses, proteins and nucleic acids. 10. The method according to claim 8, wherein part of the content of the well into which the picoliter droplet has been deposited in step 6) is extracted for a subsequent analysis and/or a scale-up of the content of the picoliter droplet by replicating a biological material. 11. The method according to claim 8, wherein in step 3), a fluorescent and/or a non-fluoresent image of the picoliter droplet is captured and processed for the sorting of the picoliter droplets. 12. The method according to claim 8, wherein in step 3), the emission of a fluorescent signal originating from a picoliter droplet is measured and is used as a selection criteria for the sorting of the picoliter droplet. 13. The method according to claim 8, wherein more than 10 picoliter droplets per second are sorted. 14. The method according to claim 8, wherein more than 100 picoliter droplets per second are sorted. 15. The method according to claim 8, wherein more than 1000 picoliter droplets per second are sorted. 16. The method according to claim 8, wherein the picoliter droplets isolated comprise a clonal population of cells in a culture medium. 17. The method according to claim 8, wherein the detection of a picoliter droplet in step 5) triggers a streaming of a gas into the detection channel via the gas inlet in order to dispense said picoliter droplet. 18. The system according to claim 1, wherein the detection channel is a glass capillary.
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이 특허에 인용된 특허 (4)
Lampert Norman R. (Norcross GA) Shevchuk George J. (Old Bridge NJ) Smith William D. (Marietta GA), Anti-snag latch assembly for a connector.
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