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US 10,544,413 B2Granted
Methods and systems for sorting droplets and beads
Assignee10x Genomics, Inc.
InventorsRajiv Bharadwaj, Michael Schnall-Levin, Anthony Makarewicz, Steven Short
Life of this patent
Estimating the term…
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.
Claims30
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;
- wherein each of said plurality of droplets comprises field-attractable particles;
- and wherein a concentration of said field-attractable particles in droplets of said second subset not having any biological particle is substantially uniform;
- (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) 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, thereby 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.
3 more elements
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;
- wherein each of said plurality of droplets comprises field-attractable particles;
- and 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;
- (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) 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, thereby 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.
3 more elements
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;
- and wherein each of said plurality of droplets comprises field-attractable particles,
- (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) 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, thereby 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;
- 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.
3 more elements
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 and 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;
- (d) 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
- (e) 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.
4 more elements
Dependencies are inferred from the claim text.