Method and Device for Encapsulating Cell in Liquid Droplet for Single-Cell Analysis

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.

1 . A method of forming droplet for single-cell analysis, the method comprising:
 providing cells;   providing one or more distinctly barcoded RNA capture beads; and   encapsulating in one droplet a single cell of the cells and a single bead of the one or more distinctly barcoded RNA capture beads,   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.   
     
     
         2 . The method of  claim 1 , 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. 
     
     
         3 . The method of  claim 1 , wherein the inertial ordering improves a ratio at which the single cell and the single bead are encapsulated in the one droplet. 
     
     
         4 . The method of  claim 1 , wherein the one or more distinctly barcoded RNA capture beads are provided with a cell lysate. 
     
     
         5 . 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. 
     
     
         6 . The method of  claim 5 , wherein the multiple nucleotides or oligonucleotides include a nucleotide sequence for capture of mRNAs in the single cell. 
     
     
         7 . The method of  claim 5 , wherein a sequence the multiple nucleotides or oligonucleotides includes an oligo-dT sequence or a primer sequence. 
     
     
         8 . A microfluidic device for forming droplet for single-cell analysis, comprising:
 a cell inlet including a first channel;   an RNA capture bead inlet including a second channel;   an oil phase inlet including a third channel;   an intersection point at which the first channel, the second channel, and the third channel are combined; and   an outlet connected to the intersection point and discharging droplet containing a cell and an RNA capture bead, wherein the first channel or the second channel is configured for a cell or an RNA capture bead in a fluid to occur inertial ordering.   
     
     
         9 . The device of  claim 8 , wherein the first channel or the second channel is spiral shaped. 
     
     
         10 . The device of  claim 8 , wherein the inertial ordering improves a ratio at which one cell and one bead are encapsulated in one droplet. 
     
     
         11 . The device of  claim 8 , wherein the one or more distinctly barcoded RNA capture beads are provided with a cell lysate. 
     
     
         12 . The device of  claim 8 , wherein each molecularly barcoded on a surface of the one or more distinctly barcoded RNA capture beads include multiple nucleotides or oligonucleotides, each molecularly barcoded on a surface of each molecularly barcoded on a surface of the one or more distinctly barcoded RNA capture beads. 
     
     
         13 . The device of  claim 12 , wherein the multiple nucleotides or oligonucleotides include a nucleotide sequence for capturing mRNA in the cells. 
     
     
         14 . The device of  claim 12 , wherein a sequence of the multiple nucleotides or oligonucleotides includes an oligo-dT sequence or a primer sequence. 
     
     
         15 . The device of  claim 8 , wherein the cell inlet, the RNA capture bead, or the oil phase inlet further includes a filter. 
     
     
         16 . The device of  claim 8 , wherein the first channel and the second channel are spiral shaped.