Precise delivery of components into fluids

Patent metadata

Field	Value
Publication number	US-11976269-B2
Application number	US-202016799464-A
Country	US
Kind code	B2
Filing date	Feb 24, 2020
Priority date	Mar 18, 2019
Publication date	May 7, 2024
Grant date	May 7, 2024

Abstract

Disclosed herein include systems, apparatuses, devices, and methods for introducing one or more components into a fluid. A first fluid and a second fluid can be co-injected into a fluidic channel of a flow cell. In some embodiments, the first fluid and a second fluid are immiscible (e.g. an aqueous buffer and a non-aqueous liquid). In some embodiments, the second fluid is less dense than the first fluid.

First claim

What is claimed is:

1. A method for introducing one or more components to contents of microwells comprising:
(a) introducing a first fluid into a fluidic channel comprising a ceiling, a first sidewall, and a bottom,
wherein the bottom of the fluidic channel comprises a plurality of microwells,
whereby the fluidic channel and each microwell of the plurality of microwells comprise the first fluid;
(b) introducing a first displacement fluid into the fluidic channel to displace the first fluid from the fluidic channel at a first flow rate;
(c) introducing a second fluid into the fluidic channel at a second flow rate while in contact with a second displacement fluid that is immiscible to the second fluid such that the second fluid is immediately followed by and in contact with the second displacement fluid and the second fluid and the second displacement fluid have the same flow rate,
wherein one or more components of the second fluid enters the content in the microwell when the second fluid comes into contact with the content in the microwell for a first duration, and
wherein the second displacement fluid displaces the second fluid from the fluidic channel and/or seals the content of the microwell.

2. The method of claim 1 , comprising:
(d) introducing a third fluid into the fluidic channel at a third flow rate while in contact with a third displacement fluid that is immiscible to the third fluid such that the third fluid is immediately followed by and in contact with the third displacement fluid and the third fluid and the third displacement fluid have the same flow rate,
wherein one or more components of the third fluid enters the content in the microwell when the third fluid comes into contact with the content in the microwell for a second duration, and
wherein the third displacement fluid displaces the third fluid from the fluidic channel and/or seals the content of the microwell.

3. The method of claim 1 , comprising introducing a third displacement fluid immediately prior to introducing a second fluid of the plurality of second fluids.

4. The method of claim 1 , comprising introducing a third displacement fluid immediately after introducing the second displacement fluid.

5. The method of claim 1 , comprising introducing a third displacement fluid immediately prior to introducing the second displacement fluid.

6. The method of claim 1 , comprising introducing a third fluid immediately prior to introducing a second fluid of the plurality of second fluids.

7. The method of claim 1 , comprising introducing a third fluid immediately after introducing the second displacement fluid.

8. The method of claim 1 , comprising introducing a third fluid immediately prior to introducing the second displacement fluid.

9. The method of claim 1 , wherein the second fluid and/or the third fluid enters the content of the microwell by diffusion.

10. The method of claim 1 , wherein the flow rate of the first fluid ranges from 0.001 ml/sec to 100 ml/sec.

11. The method of claim 10 ,
wherein the fluidic channel further comprises an anterior position and a posterior position along a longitudinal path of the fluidic channel, the anterior position being located nearer to an inlet wherein the first fluid is introduced into the fluidic channel, and
wherein the ratio of the flow rate of the first fluid at the anterior position relative the posterior position ranges from 1:1 to 1:10000.

12. The method of claim 10 ,
wherein the first duration of the contact depends on the first speed of the second fluid in the flow channel and the longitudinal length of the second fluid in the flow channel, and/or
wherein the second duration of the contact depends on the second speed of the third fluid in the flow channel and the longitudinal length of the third fluid in the flow channel.

13. The method of claim 12 ,
wherein the longitudinal length of the second fluid in the flow channel depends on the volume of the second fluid introduced, the volume of the fluidic channel, the volume of the flowcell, or a combination thereof, and/or
wherein the longitudinal length of the third fluid in the flow channel depends on the volume of the third fluid introduced, the volume of the fluidic channel, the volume of the flowcell, or a combination thereof.

14. The method of claim 1 , wherein the first flow rate is a fixed flow rate, wherein the second flow rate is a fixed flow rate, and/or wherein the third flow rate is a fixed flow rate.

15. The method of claim 1 , wherein the first flow rate is a variable flow rate, wherein the second flow rate is a variable flow rate, and/or wherein the third flow rate is a variable flow rate.

16. The method of claim 1 , wherein the first flow rate is an increasing flow rate, wherein the second flow rate is an increasing flow rate, and/or the third flow rate is an increasing flow rate.

17. The method of claim 1 , wherein the first flow rate is a decreasing flow rate, wherein the second flow rate is a decreasing flow rate, and/or wherein the third flow rate is a decreasing flow rate.

18. The method of claim 1 , wherein the second fluid and the second displacement fluid are dispensed together into the fluidic channel from a container comprising both the second fluid and the second displacement fluid.

19. The method of claim 18 , wherein the second fluid and the second displacement fluid are dispensed together into the fluidic channel from the container using a pump.

20. The method of claim 18 , wherein the container is a pipette.

21. The method of claim 20 , wherein the second fluid and the second displacement fluid are aspirated sequentially into the pipette prior to being dispensed together.

22. The method of claim 1 , wherein the fluidic channel further comprises an inlet and an outlet, and
wherein the second displacement fluid begins displacing the second fluid from the fluidic channel and/or sealing the contents of the microwell immediately adjacent to the inlet before the second fluid has reached the outlet.

Assignees

• Cellular Res Inc

Inventors

• Spuhler Philipp
• Li Sixing
• Prosen Dennis

Classifications

• C12N15/1003 Primary

  Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor · CPC title

• B01L3/5027

  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 · CPC title

• B01L2200/0673

  Handling of plugs of fluid surrounded by immiscible fluid · CPC title

• B01L2200/0642

  Filling fluids into wells by specific techniques · CPC title

• B01L2300/0816

  Cards, e.g. flat sample carriers usually with flow in two horizontal directions · CPC title