System and methods for making and processing emulsions
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12M-001/00
C12M-003/00
C12M-001/34
C12M-001/36
C12M-001/38
C12P-019/34
C12M-001/26
B01F-005/04
B01F-013/00
B01L-003/00
B01L-007/00
B04B-005/04
B04B-007/02
C12Q-001/68
출원번호
US-0775855
(2013-02-25)
등록번호
US-9121047
(2015-09-01)
발명자
/ 주소
Schultz, Jonathan
Nobile, John
Reed, Brian
Thwar, Prasanna
Roswech, Todd
Sheridan, Andrew
출원인 / 주소
Life Technologies Corporation
인용정보
피인용 횟수 :
2인용 특허 :
10
초록▼
An automated template bead preparation system is provided and includes a membrane-based emulsion generation subsystems, a thermal plate and subsystem, and a continuous centrifugation emulsion breaking and templated bead collection subsystem. The emulsion generation subsystem provides uniformity in t
An automated template bead preparation system is provided and includes a membrane-based emulsion generation subsystems, a thermal plate and subsystem, and a continuous centrifugation emulsion breaking and templated bead collection subsystem. The emulsion generation subsystem provides uniformity in the preparation of an inverse emulsion and may be used to create large or small volume inverse emulsions rapidly and reproducibly. An emulsion-generating device is provided that can supply a continuous stream of an inverse emulsion to a thermal subsystem, in automated fashion. The thermal subsystem can treat an inverse emulsion passed therethrough. The continuous centrifugation subsystem can continuously break a thermally cycled inverse emulsion and collect template beads formed in the aqueous microreactor droplets of the inverse emulsion.
대표청구항▼
1. A method of generating an amplified sample, the method comprising: generating an emulsion from a carrier fluid and a sample fluid with an emulsion generator, the emulsion generator including a carrier fluid port to receive the carrier fluid, a sample fluid port to receive the sample fluid, and an
1. A method of generating an amplified sample, the method comprising: generating an emulsion from a carrier fluid and a sample fluid with an emulsion generator, the emulsion generator including a carrier fluid port to receive the carrier fluid, a sample fluid port to receive the sample fluid, and an emulsion outlet port, the emulsion generator including a membrane to generate an emulsion including the carrier fluid and the sample fluid, the emulsion exiting the emulsion generator via the emulsion outlet port, the sample fluid including conjugated beads;heating the emulsion to an amplification condition with a heater of an amplification device, the amplification device including an inlet port in fluid communication with the emulsion outlet port and to receive the emulsion, the amplification device including an effluent port, the amplified emulsion exiting the amplification device via the effluent port, the amplified emulsion including amplified beads derived from the conjugated beads; andbreaking the emulsion following heating with a centrifuge device, the centrifuge device including an inlet conduit in fluid communication with the effluent port and to receive the emulsion after heating, the centrifuge device further including a rotor to receive a tube and a slinger to dispense the amplified emulsion received via the inlet conduit to the tube when the rotor is spinning, the amplified beads being dispensed to the tube. 2. The method of claim 1, wherein the tube includes an extension fluid passage extending from a mouth of the tube and radially outward and wherein the centrifuge further includes a peripheral gutter disposed around the rotor and to receive fluid from the extension fluid passage of the tube, wherein breaking the emulsion includes dispensing the emulsion into the slinger when the rotor is spinning. 3. The method of claim 1, wherein the centrifuge further includes an adapter to receive the inlet conduit. 4. The method of claim 3, wherein the adapter includes a casing including a central cavity and a carriage disposed within the cavity, the carriage including a bore to receive a needle coupled to a terminal end of the inlet conduit, the adapter further includes a spring to motivate the carriage away from the slinger. 5. The method of claim 1, wherein the emulsion generator further includes a first channel gasket and a second channel gasket disposed on opposite sides of the membrane, wherein generating the emulsion includes flowing the carrier fluid and the sample fluid through the membrane at least twice. 6. The method of claim 1, wherein the emulsion generator further includes a sample vial to couple to the sample fluid port and includes a displacement fluid inlet extending into a sample vial coupled to the sample fluid port. 7. An integrated apparatus comprising: an emulsion generator including a carrier fluid port to receive a carrier fluid, a sample fluid port to receive a sample fluid immiscible with the carrier fluid, and an emulsion outlet port, the emulsion generator including a membrane to generate an emulsion including the carrier fluid and the sample fluid, the emulsion exiting the emulsion generator via the emulsion outlet port;an amplification device including an inlet port in fluid communication with the emulsion outlet port and to receive the emulsion, the amplification device including a heater to subject the emulsion to amplification conditions and including an effluent port, the amplified emulsion exiting the amplification device via the effluent port; anda centrifuge device including an inlet conduit in fluid communication with the effluent port and to receive the amplified emulsion, the centrifuge device further including a rotor to receive a tube and including a slinger to dispense the amplified emulsion received via the inlet conduit to the tube when the rotor is spinning. 8. The apparatus of claim 7, wherein the tube includes an extension fluid passage extending from a mouth of the tube and radially outward. 9. The apparatus of claim 8, wherein the centrifuge further includes a peripheral gutter disposed around the rotor and to receive fluid from the extension fluid passage of the tube. 10. The apparatus of claim 7, wherein the centrifuge further includes an adapter to receive the inlet conduit. 11. The apparatus of claim 10, wherein the adapter includes a casing including a central cavity and a carriage disposed within the cavity, the carriage including a bore to receive a needle coupled to a terminal end of the inlet conduit. 12. The apparatus of claim 11, wherein the adapter further includes a spring to motivate the carriage away from the slinger. 13. The apparatus of claim 11, wherein the casing further includes a wash fluid inlet. 14. The apparatus of claim 7, wherein the emulsion generator further includes a first channel gasket and a second channel gasket disposed on opposite sides of the membrane and defining a fluid path passing through the membrane at least twice. 15. The apparatus of claim 7, wherein the fluid path is concurrent. 16. The apparatus of claim 7, wherein the fluid path is countercurrent. 17. The apparatus of claim 7, wherein the emulsion generator further includes a sample vial to couple to the sample fluid port. 18. The apparatus of claim 17, wherein the emulsion generator further includes a displacement fluid inlet extending into a sample vial coupled to the sample fluid port. 19. The apparatus of claim 7, wherein the amplification device further includes an amplification plate to receive the emulsion from the emulsion outlet port. 20. The apparatus of claim 19, wherein the heater is to cycle the temperature of the amplification plate.
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