Microfluidic reporter cell assay methods and kits thereof

Abstract

Functional assays using reporter cell assays are described which probe the activity of at least one cell of interest. The ability to probe at least one cell is provided by using the microfluidic methods, devices and kits described herein. Assays combining both reporter cell signaling as well as binding assay signaling for at least one cell is also described herein.

1 .- 67 . (canceled) 
     
     
         68 . A method of assaying at least one biological cell for a biological activity in a microfluidic device comprising:
 an enclosure having an inner lower surface and an inner upper surface spaced apart defining a chamber, comprising a flow region configured to contain a flow of a first fluidic medium, and at least one incubation chamber, each of which is disposed upon the inner lower surface,   wherein the at least one incubation chamber is enclosed by walls extending from the inner lower surface to the inner upper surface of the chamber and has a single lateral opening to the flow region, and   wherein the single lateral opening between the flow region and the connection region is configured to facilitate substantially only diffusion between the first fluidic medium and the second fluidic medium in the isolation region when the first fluidic medium is flowing through the flow region, the method comprising:
 introducing the at least one biological cell into the at least one incubation chamber; 
 introducing at least one reporter cell into the at least one incubation chamber, wherein the at least one reporter cell is configured to produce a first detectable signal when the at least one biological cell comprises the biological activity; and 
 analyzing the at least one reporter cell for an activity stimulated by the presence of a biological activity of the at least one biological cell. 
   
     
     
         69 . The method of  claim 68 , wherein analyzing comprises incubating the at least one biological cell and the one at least one reporter cell in the at least one incubation chamber for a pre-determined period of time, thereby allowing the one at least one reporter cell to produce the first detectable signal. 
     
     
         70 . The method of  claim 69 , wherein incubating further comprises providing the one at least one reporter cell with one or more reagents forming one or all of the components of the detectable signal of the at least one reporter cell. 
     
     
         71 . The method of  claim 68 , wherein analyzing further comprises providing excitation light to excite a fluorophore of the first detectable signal of the at least one reporter cell. 
     
     
         72 . The method of  claim 71 , further comprising detecting the excited fluorophore. 
     
     
         73 . The method of  claim 68 , further comprising introducing at least one capture micro-object into at least the flow region. 
     
     
         74 . The method of  claim 73 , further comprising introducing one or more visualization reagents which are configured to bind to the at least one capture micro-object to produce a second detectable signal. 
     
     
         75 . The method of  claim 74 , further comprising detecting the second detectable signal. 
     
     
         76 . The method of  claim 68 , wherein introducing the at least one biological cell into the at least one incubation chamber of the microfluidic device comprises using a DEP force having sufficient strength to move the at least one biological cell. 
     
     
         77 . The method of  claim 76 , further comprising optically actuating the DEP force. 
     
     
         78 . The method of  claim 68 , wherein the at least one biological cell is a mammalian cell. 
     
     
         79 . The method of  claim 68 , wherein the at least one biological cell is a hybridoma. 
     
     
         80 . The method of  claim 68 , wherein the at least one biological cell is a lymphocyte or a leukocyte. 
     
     
         81 . The method of  claim 68 , wherein the at least one incubation chamber further comprises:
 an isolation region having a single opening and configured to contain a second fluidic medium; and   a connection region, comprising a distal opening to the isolation region and a proximal opening comprising the single lateral opening into the flow region.   
     
     
         82 . The method of  claim 81 , further comprising introducing the at least one reporter cell into an isolation region of the at least one incubation chamber. 
     
     
         83 . The method of  claim 82 , further comprising introducing the at least one biological cell into the isolation region of the at least one incubation chamber. 
     
     
         84 . The method of  claim 68 , wherein the single lateral opening into the microfluidic channel of the at least one incubation chamber has a width ranging from about 20 microns to about 100 microns. 
     
     
         85 . The method of  claim 68 , wherein a distance between the inner lower surface and the inner upper surface of the enclosure defining the chamber is from about 30 to about 200 microns. 
     
     
         86 . The method of  claim 68 , wherein a distance between the inner lower surface and the inner upper surface of the enclosure defining the chamber is a substantially uniform distance. 
     
     
         87 . The method of  claim 68 , wherein the at least one incubation chamber further comprises at least one surface conditioned to support cell growth, viability, portability, or any combination thereof.