Cellular cassettes for the collection, storage, and analysis of biological samples

Abstract

Disclosed herein are cellular cassettes for the storage, collection, and analysis of biological samples. The cellular cassette can enable easy sample collection and sealing of microwell arrays with semi-permeable membrane for stable storage and future processing of single cells. Also disclosed herein are systems and kits comprising one or more described cassettes. The described cassettes, systems, and kits can be used to create barcoded, single-cell sequencing libraries. Further described herein are methods of using the cassettes, systems, and kits.

1 .- 149 . (canceled) 
     
     
         150 . A cassette for collecting, storing, or analyzing bioparticles, said cassette comprises:
 (a) a loading assembly that comprises a microwell array and a locking member,
 wherein said microwell array comprises a plurality of microwells situated on a base and a plurality of beads; and 
   (b) a loading ring reversibly attached to the base, wherein the loading ring is configured to retain excess fluid on the microwell array.   
     
     
         151 . The cassette of  claim 150 , wherein the loading ring is configured to retain from about 0.1 ml to about 5 ml fluid. 
     
     
         152 . The cassette of  claim 150 , wherein the loading ring is configured to retain from about 0.5 to 1.5 ml fluid. 
     
     
         153 . The cassette of  claim 150 , wherein the loading assembly further comprises an injection lid that covers at least a portion of the microwell array. 
     
     
         154 . The cassette of  claim 153 , wherein the injection lid covers at least 80% of the microwell array. 
     
     
         155 . The cassette of  claim 153 , wherein the injection lid has an opening that is configured to direct pipetting. 
     
     
         156 . The cassette of  claim 150 , wherein the microwell array comprises at least 1000 microwells that each comprises a single cell. 
     
     
         157 . The cassette of  claim 150 , wherein the loading assembly further comprises a port for fluid collection. 
     
     
         158 . The cassette of  claim 150 , wherein the loading assembly further comprises a key hole. 
     
     
         159 . The cassette of  claim 150 , wherein the microwell array comprises a wetting solution or a wash buffer. 
     
     
         160 . A method for collecting, storing, or analyzing bioparticles using a cellular cassette, wherein said cellular cassette comprises:
 (a) a loading assembly that comprises a microwell array and a locking member,
 wherein said microwell array comprises a plurality of microwells situated on a base and a plurality of beads; and 
   (b) a loading ring reversibly attached to the base, wherein the loading ring is configured to retain excess fluid on the microwell array;   wherein said method comprises   (a) loading a sample fluid that comprises one or more cells onto said microwell array, thereby loading at least one cell into one of said microwells.   
     
     
         161 . The method of  claim 160 , further comprising wetting the microwell array before the sample fluid is loaded. 
     
     
         162 . The method of  claim 160 , further comprising mixing the loaded sample fluid. 
     
     
         163 . The method of  claim 160 , further comprising incubating the loaded sample fluid from about 30 seconds to about 12 hours. 
     
     
         164 . The method of  claim 160 , further comprising applying a storage buffer to the microwell array after the sample fluid is loaded, wherein the storage buffer is configured to preserve polynucleic acids. 
     
     
         165 . The method of  claim 160 , further comprising removing the loading ring after the sample fluid is loaded. 
     
     
         166 . The method of  claim 165 , further comprising applying a membrane assembly onto the loading assembly, wherein the membrane assembly comprises
 (a) a semi-permeable membrane that comprises a membrane top surface and a membrane bottom surface, and   (b) a membrane frame configured to reversibly attach to said loading assembly, wherein said semi-permeable membrane is attached to said membrane frame.   
     
     
         167 . The method of  claim 166 , wherein the membrane assembly is reversibly attached to a plunger assembly that comprises a plunger, and wherein said plunger comprises a bottom surface. 
     
     
         168 . The method of  claim 167 , wherein the membrane assembly is pre-assembled with the plunger assembly. 
     
     
         169 . The method of  claim 167 , wherein the plunger assembly or the membrane assembly comprises a top locking member that is configured to engage with said locking member of the loading assembly.