Cell sorting system and methods

Abstract

Apparatus and Methods are provided for a microfabricated fluorescence activated cell sorter based on a switch for rapid, active control of cell routing through a microfluidic channel network. This sorter enables low-stress, highly efficient sorting of populations of small numbers of cells (i.e., 1000 - 100,000 cells). The invention includes packaging of the microfluidic channel network in a self-contained plastic cartridge that enables microfluidic channel network to macro-scale instrument interconnect, in a sterile, disposable format. Optical and/or fluidic switching forces are used alone or in combination to effect switching.

What is claimed: 
     
         1 - 24 . (canceled) 
     
     
         25 . An integrated structure for microfluidic analysis and sorting comprising:
 a cartridge, the cartridge comprising
 an optical window, 
 a plurality of reservoirs, including at least:
 a sample reservoir, 
 a fluid reservoir, 
 a waste reservoir, and 
 a target collection reservoir, 
 
   a chip, the chip including at least:
 a sample inlet channel, the cell inlet channel being fluidically coupled to the sample reservoir, 
 one or more fluid inlet channels, the fluid inlet channels being fluidically coupled to the fluid reservoir, 
 a detection region, 
 a branched sorting region, and 
 at least two outlet channels including at least a waste channel and a target channel, waste channel being fluidically coupled to the waste reservoir and the target channel being coupled to the target collection reservoir, 
 the chip being disposed adjacent the optical window, 
   a lid, the lid including at least:
 a pneumatic pressure port, the port having an inlet and being coupled to at least one of the sample reservoir and the fluid reservoir, and 
 a filter disposed between the inlet of the pneumatic pressure port and at least one of the sample reservoir and the fluid reservoir, and 
 a manifold, the manifold coupling pneumatic pressure from a source to the pneumatic pressure port without intervening tubing. 
   
     
     
         26 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the sample reservoir is conical. 
     
     
         27 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the sample reservoir is tapered. 
     
     
         28 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the sample reservoir further includes an insert. 
     
     
         29 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the insert is a polypropylene insert. 
     
     
         30 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the lid is plastic. 
     
     
         31 . The integrated structure for microfluidic analysis and sorting of  claim 30  wherein the lid is acrylic plastic. 
     
     
         32 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the lid includes separate pneumatic pressure ports for the sample reservoir and the fluid reservoir. 
     
     
         33 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the lid includes a pneumatic pressure port for the waste reservoir. 
     
     
         34 . The integrated structure for microfluidic analysis and sorting of  claim 33  wherein the lid further includes a pneumatic pressure port for the target collection reservoir. 
     
     
         35 . The integrated structure for microfluidic analysis and sorting of  claim 32  wherein the lid includes a pneumatic pressure port for the waste reservoir. 
     
     
         36 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the lid provides optical access to the reservoirs. 
     
     
         37 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the lid includes snap-on attachments to couple to the cartridge. 
     
     
         38 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the filter is gas permeable. 
     
     
         39 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the filter is fluid impermeable. 
     
     
         40 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the filter is a polypropylene filter. 
     
     
         41 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein curable adhesive is disposed between the chip and the cartridge. 
     
     
         42 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein the adhesive is UV curable. 
     
     
         43 . The integrated structure for microfluidic analysis and sorting of  claim 25  further including a bonding sheet between the chip and the cartridge. 
     
     
         44 . (canceled) 
     
     
         45 . The integrated structure for microfluidic analysis and sorting of  claim 25  wherein optical access to the chip is provided through he optical window and through the reverse side of the chip. 
     
     
         46 - 51 . (canceled)