Methods and systems for sorting droplets and beads

Abstract

Methods and systems for sorting droplets are provided. In some cases, occupied droplets may be sorted from unoccupied droplets. In some cases, singularly occupied droplets may be sorted from unoccupied droplets and multiply occupied droplets. Methods and systems for sorting cell beads are provided. In some cases, cell beads may be sorted from particles unoccupied with cell derivatives. In some cases, singularly occupied cell beads may be sorted from unoccupied particles and multiply occupied cell beads. Methods and systems for selectively polymerizing droplets based on occupancy and size of the droplets are provided.

What is claimed is: 
     
         1 . A method for sorting droplets, comprising:
 (a) bringing a first phase in contact with a second phase immiscible with said first phase, to generate a plurality of droplets, wherein said plurality of droplets comprises (i) a first subset of droplets each including, and not more than, one biological particle, and (ii) a second subset of droplets each either having more than one biological particle or not having any biological particle, wherein said biological particle is a cell, a derivative of a cell, or a constituent of a cell;   (b) directing said plurality of droplets along a first channel towards an intersection of said first channel with at least a second channel and a third channel; and   (c) separating at least a portion of said first subset of said plurality of droplets from at least a portion of said second subset of said plurality of droplets, wherein upon separation, said at least said portion of said first subset of said plurality of droplets flows along said second channel and said at least said portion of said second subset of said plurality of droplets flows along said third channel.   
     
     
         2 . The method of  claim 1 , wherein said first subset of said plurality of droplets includes particles having coupled thereto molecules comprising barcode sequences. 
     
     
         3 . The method of  claim 2 , wherein said particles are gel beads. 
     
     
         4 . The method of  claim 1 , further comprising detecting individual droplets of said first subset of said plurality of droplets, and subjecting said individual droplets to a stimulus to facilitate polymerization in said biological particles upon detecting said individual droplets. 
     
     
         5 . The method of  claim 4  wherein said stimulus is applied prior to said intersection. 
     
     
         6 . The method of  claim 4 , wherein said stimulus is applied subsequent to said intersection. 
     
     
         7 . The method of  claim 4 , wherein said stimulus is an optical stimulus or chemical stimulus. 
     
     
         8 . The method of  claim 1 , wherein said biological particles are cells enclosed within or comprising a gel or polymer matrix. 
     
     
         9 . The method of  claim 1 , wherein each of said plurality of droplets comprises field-attractable particles, and wherein (c) comprises subjecting said plurality of droplets to an electric or magnetic field under conditions sufficient to separate said at least said portion of said first subset from said at least said portion of said second subset. 
     
     
         10 . The method of  claim 9 , wherein a concentration of said field-attractable particles in droplets of said second subset not having any biological particle is substantially uniform. 
     
     
         11 . The method of  claim 9 , wherein each droplet of said first subset comprises (i) less field attractable particles than each droplet of said second subset not having any biological particle, and (ii) more field attractable particles than each droplet of said second subset having more than one biological particle. 
     
     
         12 . The method of  claim 11 , wherein forces induced by said electric or magnetic field on droplets of said second subset not having any biological particle are greater than forces induced on said first subset, and wherein forces induced on said first subset are greater than forces induced on droplets of said second subset having more than one biological particle. 
     
     
         13 . The method of  claim 9 , wherein said field-attractable particles are magnetic-field attractable particles. 
     
     
         14 . The method of  claim 9 , wherein said conditions of said electric or magnetic field sufficient to separate said at least said portion of said first subset and said at least said portion of said second subset are determined based at least in part on a ratio between sizes of said plurality of droplets and sizes of said biological particles in said first subset of said plurality of droplets. 
     
     
         15 . The method of  claim 1 , further comprising, subsequent to (c), subjecting nucleic acid molecules derived from said biological particles in said first subset to nucleic acid sequencing. 
     
     
         16 . The method of  claim 15 , further comprising, subsequent to (c), subjecting said first subset of said plurality of droplets to conditions sufficient to yield extension products of said nucleic acid molecules from said biological particles in said first subset, and subjecting said extension products or derivatives thereof to nucleic acid sequencing. 
     
     
         17 . The method of  claim 1 , wherein (c) comprises subjecting said plurality of droplets to a pressure pulse under conditions sufficient to separate said at least said portion of said first subset from said at least said portion of said second subset. 
     
     
         18 . The method of  claim 17 , wherein forces induced by said pressure pulse on droplets of said second subset not having any biological particle are greater than forces induced on said first subset, and wherein forces induced on said first subset are greater than forces induced on droplets of said second subset having more than one biological particle. 
     
     
         19 . The method of  claim 1 , wherein upon separation, said at least said portion of said first subset and droplets of said second subset having more than one biological particle flow along said second channel and droplets of said second subset not having any biological particle flow along said third channel. 
     
     
         20 . The method of  claim 19 , further comprising (i) directing said at least said portion of said first subset and said droplets of said second subset having more than one biological particle along said second channel towards a second intersection of said second channel with at least a fourth channel and a fifth channel, and (ii) separating said at least said portion of said first subset from at least a portion of said droplets of said second subset having more than one biological particle, wherein upon separation, said at least said portion of said first subset flows along said fourth channel and said at least said portion of said droplets of said second subset having more than one biological particle flows along said fifth channel. 
     
     
         21 . A method for sorting particles, comprising:
 (a) providing a plurality of particles comprising (i) a first set of particles having a plurality of cells or material therefrom, and (ii) a second set of particles each not having any cell or material therefrom; and   (b) sorting said plurality of particles, thereby isolating at least a portion of said first set of particles from at least a portion of said second set of particles.   
     
     
         22 . The method of  claim 21 , wherein said first set of particles comprises (i) a first subset of particles each including, but not more than, one cell of said plurality of cells, or material from said cell, and (ii) a second subset of particles each including more than one cell of said plurality of cells, or material from said more than one cell. 
     
     
         23 . The method of  claim 22 , further comprising isolating at least a portion of said first subset of particles from at least a portion of said second subset of particles. 
     
     
         24 . The method of  claim 21 , wherein (b) comprises subjecting said plurality of particles to a magnetic or electric field to isolate said at least said portion of said first subset of particles from said at least said portion of said second subset of particles. 
     
     
         25 . The method of  claim 24 , wherein each particle of said plurality of particles comprises field-attractable particles. 
     
     
         26 . The method of  claim 21 , wherein (b) comprises subjecting said plurality of particles to a pressure pulse to isolate said at least said portion of said first subset of particles from said at least said portion of said second subset of particles. 
     
     
         27 . The method of  claim 21 , wherein each of said plurality of particles comprises a gel or polymer matrix. 
     
     
         28 . The method of  claim 27 , further comprising, prior to (a), generating said plurality of particles by (i) encapsulating each of said plurality of cells or material therefrom in a gel or polymer matrix, or (ii) generating said gel or polymer matrix in each of said plurality of cells. 
     
     
         29 . The method of  claim 21 , further comprising, subsequent to (b), subjecting nucleic acid molecules derived from said first subset to nucleic acid sequencing. 
     
     
         30 . The method of  claim 21 , wherein a particle of said first set of particles comprises a cell of said plurality of cells, or material derived from said cell.