Microfluidic Devices and Methods for Use Thereof in Multicellular Assays of Secretion

Abstract

Methods and devices are provided herein for identifying a cell population comprising an effector cell that exerts an extracellular effect. In one embodiment the method comprises retaining in a microreactor a cell population comprising one or more effector cells, wherein the contents of the microreactor further comprise a readout particle population comprising one or more readout particles, incubating the cell population and the readout particle population within the microreactor, assaying the cell population for the presence of the extracellular effect, wherein the readout particle population or subpopulation thereof provides a direct or indirect readout of the extracellular effect, and determining, based on the results of the assaying step, whether one or more effector cells within the cell population exerts the extracellular effect on the readout particle. If an extracellular effect is measured, the cell population is recovered for further analysis to determine the cell or cells responsible for the effect.

1 - 218 . (canceled) 
     
     
         219 . A method of identifying an antibody secreting cell (ASC) that produces a virus-neutralizing antibody, comprising:
 retaining in a plurality of microreactors a plurality of cell populations, wherein individual cell populations of the plurality of cell populations comprise one or more ASCs and are retained in individual microreactors of the plurality of microreactors, wherein the individual microreactors of the plurality of microreactors each comprise one or more readout cells;   introducing a plurality of accessory particle populations into the plurality of microreactors, wherein the plurality of accessory particle populations comprise a plurality of virus particles operable to infect the one or more readout cells and individual accessory particle populations of the plurality of accessory particle populations are retained in the individual microreactors;   incubating the individual cell populations, the readout cells, and the accessory particle populations within the individual microreactors for a time sufficient to produce a plurality of antibodies;   assaying the individual microreactors and determining whether the plurality of antibodies comprises a virus-neutralizing antibody; and   identifying the antibody secreting cell (ASC) that produces the virus-neutralizing antibody.   
     
     
         220 . The method of  claim 219 , wherein the plurality of virus particles is engineered to include a fluorescent protein expressed by the one or more readout cells following virus infection of the one or more readout cells. 
     
     
         221 . The method of  claim 220 , wherein assaying the individual microreactors comprises assaying the fluorescent protein expressed by the one or more readout cells following virus infection of the one or more readout cells. 
     
     
         222 . The method of  claim 219 , wherein assaying the individual microreactors comprises assaying the death of the one or more readout cells following virus infection of the one or more readout cells. 
     
     
         223 . The method of  claim 219 , further comprising substantially isolating the individual microreactors from their surrounding environments. 
     
     
         224 . The method of  claim 220 , further comprising substantially isolating the individual microreactors from their surrounding environments. 
     
     
         225 . The method of  claim 221 , further comprising substantially isolating the individual microreactors from their surrounding environments. 
     
     
         226 . The method of  claim 219 , wherein if an individual cell population comprises an ASC that produces a virus-neutralizing antibody, the method further comprises a recovering step comprising recovering the individual cell population comprising the ASC that produces the virus-neutralizing antibody or a portion thereof to obtain a recovered cell population. 
     
     
         227 . The method of  claim 226 , wherein the recovering step comprises positioning the open end of a microcapillary in a microreactor comprising the cell population comprising the ASC that produces the virus neutralizing antibody and aspirating contents of the microreactor or a portion thereof to obtain a recovered aspirated cell population. 
     
     
         228 . The method of  claim 227 , wherein the microcapillary is mounted on a robotic micromanipulation system on a microscope or the microcapillary is controlled robotically. 
     
     
         229 . The method of  claim 226 , further comprising,
 retaining a plurality of cell subpopulations originating from the recovered cell population in a plurality of vessels, wherein each cell subpopulation is present in an individual vessel,   lysing individual cell subpopulations of the plurality of cell subpopulations to provide lysed cell subpopulations, and   amplifying one or more nucleic acids within each of the lysed cell subpopulations.