Amplifying rare cell surface markers

Abstract

This invention relates generally to a microfluidic device for encapsulation, incubation, and analysis of cell surface markers or secreted molecules from a single cell.

1 - 8 . (canceled) 
     
     
         9 . A system for detecting expression of a specific protein or peptide by individual cells of a population of cells, the system comprising:
 a fluid control device configured to divide a sample comprising the population of cells into subsamples, such that each subsample contains at most one of the population of cells and each subsample is encompassed a hydrophobic fluid;   rolling circle amplification reagents for detection of the presence of the specific protein or peptide; and   a device operable to detect the rolling circle amplification reagents in individual subsamples.   
     
     
         10 . The system of  claim 9 , wherein the rolling circle amplification reagents comprise a phi29 DNA polymerase or a circular DNA template or both. 
     
     
         11 . The system of  claim 9 , wherein the rolling circle amplification reagents comprise a biotinylated antibody that binds specifically to the protein or peptide, an avidin bridge, a biotinylated DNA probe, and a circular DNA template capable of hybridizing with the biotinylated DNA probe. 
     
     
         12 . The system of  claim 11 , wherein the rolling circle amplification reagents further comprise a phi29 polymerase and a fluorescently-labeled nucleotide. 
     
     
         13 . The system of  claim 11 , wherein the biotinylated antibody binds specifically to a protein or peptide expressed by a cancer cell, a stem cell, or an immune cell. 
     
     
         14 . The system of  claim 11 , wherein the biotinylated antibody binds specifically to an epithelial cell adhesion molecule (EpCAM). 
     
     
         15 . The system of  claim 9 , further comprising a cell sorting mechanism operable to sort the subsamples based on measurement of an indicator parameter in each subsample. 
     
     
         16 . The system of  claim 15 , wherein the cell sorting mechanism comprises a fluorescence activated cell sorting mechanism. 
     
     
         17 . The system of  claim 9 , wherein dividing the sample into subsamples comprises combining aqueous sample fluid from a first inlet channel with a hydrophobic carrier fluid. 
     
     
         18 . A kit comprising:
 a fluid control device configured to divide a sample comprising the population of cells into subsamples, such that each subsample contains at most one of the population of cells and each subsample is encompassed a hydrophobic fluid; and   rolling circle amplification reagents for detection of the presence of the specific protein or peptide.   
     
     
         19 . The kit of  claim 18 , wherein the rolling circle amplification reagents comprise a phi29 DNA polymerase or a circular DNA template or both. 
     
     
         20 . The kit of  claim 18 , wherein the rolling circle amplification reagents comprise a biotinylated antibody that binds specifically to the protein or peptide, an avidin bridge, a biotinylated DNA probe, and a circular DNA template capable of hybridizing with the biotinylated DNA probe. 
     
     
         21 . The kit of  claim 20 , wherein the rolling circle amplification reagents further comprise a phi29 polymerase and a fluorescently-labeled nucleotide. 
     
     
         22 . The kit of  claim 20 , wherein the biotinylated antibody binds specifically to a protein or peptide expressed by a cancer cell, a stem cell, or an immune cell. 
     
     
         23 . The kit of  claim 20 , wherein the biotinylated antibody binds specifically to an epithelial cell adhesion molecule (EpCAM).