Determination of cells using amplification

Abstract

The present invention generally relates to microfluidics and, in particular, to systems and methods for determining cells using amplification. In one set of embodiments, cells are encapsulated within droplets and nucleic acids from the cells amplified within the droplets. The droplets may then be pooled together and the amplified nucleic acids can be determined using PCR or other suitable techniques. In some embodiments, techniques such as these can be used to detect relatively rare cells that may be present, e.g., if the droplets are amplified using conditions able to selectively amplify nucleic acids arising from the relatively rare cells.

1 - 50 . (canceled) 
     
     
         51 . A method, comprising:
 within a plurality of droplets encapsulating nucleic acids arising from a plurality of cells, applying conditions able to selectively amplify a target nucleic acid sequence suspected of being present within the nucleic acids;   combining the interiors of the droplets together to form a combined fluid; and   determining amplified nucleic acids contained within the combined fluid.   
     
     
         52 . A method, comprising:
 encapsulating cells within a plurality of droplets;   lysing at least some of the cells within the plurality of droplets to release nucleic acids from the cells into the interior of the droplets;   within the interior of the droplets, applying conditions able to selectively amplify a target nucleic acid sequence suspected of being present within the nucleic acids released from the cells; and   determining amplified nucleic acids contained within the droplets.   
     
     
         53 . The method of  claim 51 , wherein the target nucleic acid sequence is a DNA sequence. 
     
     
         54 . The method of  claim 51 , wherein the target nucleic acid sequence is an RNA sequence. 
     
     
         55 . The method of  claim 51 , wherein the target nucleic acid sequence is an mRNA sequence. 
     
     
         56 . The method of  claim 51 , wherein applying conditions able to selectively amplify a target nucleic acid sequence comprises applying conditions able to cause amplification via PCR. 
     
     
         57 . The method of  claim 56 , wherein applying conditions able to selectively amplify a target nucleic acid sequence comprises applying conditions able to cause amplification via RT-PCR. 
     
     
         58 . The method of  claim 51 , wherein applying conditions able to selectively amplify a target nucleic acid sequence comprises using PCR to amplify the target nucleic acid sequence. 
     
     
         59 . The method of  claim 51 , wherein applying conditions able to selectively amplify a target nucleic acid sequence comprises adding a polymerase to at least some of the interior of the droplets. 
     
     
         60 . The method of  claim 59 , comprising adding a polymerase to at least some of the interior of the droplets via injection. 
     
     
         61 . The method of  claim 52 , wherein the target nucleic acid sequence is a DNA sequence. 
     
     
         62 . The method of  claim 52 , wherein the target nucleic acid sequence is an RNA sequence. 
     
     
         63 . The method of  claim 52 , wherein the target nucleic acid sequence is an mRNA sequence. 
     
     
         64 . The method of  claim 52 , wherein applying conditions able to selectively amplify a target nucleic acid sequence comprises applying conditions able to cause amplification via PCR. 
     
     
         65 . The method of  claim 64 , wherein applying conditions able to selectively amplify a target nucleic acid sequence comprises applying conditions able to cause amplification via RT-PCR. 
     
     
         66 . The method of  claim 52 , wherein applying conditions able to selectively amplify a target nucleic acid sequence comprises using PCR to amplify the target nucleic acid sequence. 
     
     
         67 . The method of  claim 52 , wherein applying conditions able to selectively amplify a target nucleic acid sequence comprises adding a polymerase to at least some of the interior of the droplets. 
     
     
         68 . The method of  claim 67 , comprising adding a polymerase to at least some of the interior of the droplets via injection.