Method and device for encapsulating cell in liquid droplet for single-cell analysis

Patent metadata

Field	Value
Publication number	US-12226773-B2
Application number	US-202217962752-A
Country	US
Kind code	B2
Filing date	Oct 10, 2022
Priority date	May 17, 2017
Publication date	Feb 18, 2025
Grant date	Feb 18, 2025

Abstract

Provided are a method and a device for encapsulating a cell in droplet for single-cell analysis, or a method and a device for forming droplet for single-cell analysis. According to the method and the device of one aspect, by using the effects of inertial ordering, not only a ratio at which one cell is encapsulated in one droplet is increased, but also a yield of generating droplet is improved.

First claim

The invention claimed is:

1. A method of forming droplets for single-cell analysis, the method comprising:
providing cells;
providing one or more distinctly barcoded RNA capture beads;
providing one or more oil phase;
encapsulating in one droplet a single cell of the cells and a single bead of the one or more distinctly barcoded RNA capture beads; and
mixing the cells and beads encapsulated in the droplet,
wherein the providing of the cells or the providing of the one or more distinctly barcoded RNA capture beads is provided through inertial ordering before the single cell or the single bead are encapsulated in the one droplet, and
wherein the inertial ordering occurs through a spiral channel of a microfluidic device to which the cells or the one or more distinctly barcoded RNA capture beads are provided,
wherein the oil phase in the providing one or more oil phase step has a flow rate in a range of 4.0 mL/h to 8.0 mL/h, and
wherein the cells in the providing cells step and beads in the providing one or more distinctly barcoded RNA capture beads step have a flow rate in a range of 8.0 mL/h to 14 mL/h 6.0 mL/h.

2. The method of claim 1 , wherein the one or more distinctly barcoded RNA capture beads are provided with a cell lysate.

3. The method of claim 1 , wherein the one or more distinctly barcoded RNA capture beads include multiple nucleotides or oligonucleotides, each molecularly barcoded on a surface of the one or more distinctly barcoded RNA capture beads.

4. The method of claim 3 , wherein the multiple nucleotides or oligonucleotides include a nucleotide sequence for capture of mRNAs in the single cell.

5. The method of claim 3 , wherein a sequence the multiple nucleotides or oligonucleotides includes an oligo-dT sequence or a primer sequence.

Assignees

• Samsung Life Public Welfare Foundation

Inventors

• Park Woong-Yang
• Moon Hui-Sung
• Kim Shin-Hyun
• Je Kwanghwi

Classifications

• C12Q1/6869 Primary

  Methods for sequencing · CPC title

• B01L2300/0867

  Multiple inlets and one sample wells, e.g. mixing, dilution · CPC title

• B01L2200/0652

  Sorting or classification of particles or molecules · CPC title

• B01L2200/0636

  Focussing flows, e.g. to laminate flows · CPC title

• B01L3/502784 Primary

  specially adapted for droplet or plug flow, e.g. digital microfluidics · CPC title