Methods for sorting particles
Methods and systems for sorting particles are provided. 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.
1. A method for identifying biological particles of interest comprising:
(a) bringing a first phase in contact with a second phase immiscible with the first phase to generate a plurality of droplets, wherein the plurality of droplets comprises (i) a first subset of droplets each including a single biological particle, and (ii) a second subset of droplets each either having more than one biological particle or not having any biological particle, wherein the biological particle is a cell or an organelle; wherein each of the plurality of droplets comprises a bead comprising field-attractable particles;
(b) directing the plurality of droplets along a first channel towards an intersection of the first channel with at least a second channel and a third channel;
(c) subjecting the plurality of droplets to an electric or magnetic field to separate a portion of the first subset of the plurality of droplets from a portion of the second subset of the plurality of droplets, wherein upon separation, the portion of the first subset of the plurality of droplets flows along the second channel and the portion of the second subset of the plurality of droplets flows along the third channel; and
(d) sequencing nucleic acid molecules derived from the biological particles from the portion of the first subset of the plurality droplets.
2. The method of claim 1 , wherein step (d) further comprises amplifying the nucleic acid molecules prior to sequencing.
3. The method of claim 2 , wherein prior to the amplifying the biological particles are lysed.
4. The method of claim 3 , wherein the biological particles are lysed in the presence of one of the beads.
5. The method of claim 4 , wherein the one of the beads is a gel bead.
6. The method of claim 4 , wherein the one of the beads comprises additional reagents for amplification and/or sequencing.
7. The method of claim 1 , wherein the field attractable particles are magnetic particles or fluorescent particles.
8. The method of claim 1 , further comprising subjecting the first subset of droplets to a second electric field or magnetic field.
9. The method of claim 1 , wherein step (c) further comprises detecting the droplets.
10. The method of claim 7 , wherein the magnetic particles are paramagnetic particles.
11. A method for identifying biological particles of interest comprising:
(a) bringing a first phase in contact with a second phase immiscible with the first phase to generate a plurality of droplets, wherein the plurality of droplets comprises (i) a first subset of droplets each including more than one biological particle, and (ii) a second subset of droplets each either having a single biological particle or not having any biological particle, wherein the biological particle is a cell or an organelle;
(b) directing the plurality of droplets along a first channel towards an intersection of the first channel with at least a second channel and a third channel;
(c) subjecting the plurality of droplets to an electric or magnetic field to separate a portion of the first subset of the plurality of droplets from a portion of the second subset of the plurality of droplets, wherein upon separation, the portion of the first subset of the plurality of droplets flows along the second channel and the portion of the second subset of the plurality of droplets flows along the third channel;
(d) lysing the biological particles in the presence of a bead; and
(e) sequencing nucleic acid molecules derived from the biological particles from the portion of the first subset of the plurality of droplets.
12. The method of claim 11 , wherein step (e) further comprises amplifying the nucleic acid molecules prior to sequencing.
13. The method of claim 12 , wherein the cells are lysed prior to the amplifying the biological particles.
14. The method of claim 11 , wherein the bead is a gel bead.
15. The method of claim 11 , wherein the bead comprises additional reagents for amplification and/or sequencing.
16. The method of claim 11 , wherein step (c) further comprises detecting the droplets.