Methods and devices for analysis of defined multicellular combinations

Abstract

Methods for cell analysis are provided, comprising cell capturing, characterization, transport, and culture. In an exemplary method individual cells (and/or cellular units) are flowed into a microfluidic channel, the channel is partitioned into a plurality of contiguous segments, capturing at least one cell in at least one segment. A characteristic of one or more captured cells is determined and the cell(s) and combinations of cells are transported to specified cell holding chamber(s) based on the determined characteristic(s). Also provided are devices and systems for cell analysis.

1 .- 52 . (canceled) 
     
     
         53 . A chamber in a microfluidic device that is configured for receiving and maintaining live cells, the chamber comprising:
 an input channel configured so that nucleated eukaryotic cells can pass intact through the input channel and into the chamber;   a plurality of four or more drain channels configured so that fluid but not nucleated eukaryotic cells can exit the chamber; and   a gas permeable membrane separating the chamber from a supply channel, wherein the supply channel is configured to bring a gaseous mixture to the chamber, and the gas permeable membrane is configured so that gasses may be exchanged between the chamber and the supply channel.   
     
     
         54 . The chamber of  claim 53 , wherein the chamber or the input channel is connected to a reagent channel configured for supplying a fluid comprising one or more reagents for maintaining or treating cells in the chamber. 
     
     
         55 . The chamber of  claim 54 , wherein the reagent channel is connected to a supply of a fluid containing cell nutrients. 
     
     
         56 . The chamber of  claim 54 , wherein the reagent channel is connected to a supply of a fluid that lyses cells upon delivery of the fluid into the chamber. 
     
     
         57 . The chamber of  claim 53 , wherein the input channel tapers towards the chamber, thereby directing cells towards the chamber's center. 
     
     
         58 . The chamber of  claim 53 , wherein the drain channels are distributed around the chamber in a manner that allows fluid to flow from the input channel into the chamber and out the drain channels without drawing cells in the chamber towards the drain channels. 
     
     
         59 . The chamber of  claim 58 , wherein the chamber has a perimeter that is substantially circular or oval in shape, and the drain channels are distributed over more than 180 degrees of the perimeter. 
     
     
         60 . (canceled) 
     
     
         61 . The chamber of  claim 53 , wherein the drain channels connect to the chamber through drain openings and the input channel connects to the channel though an input opening, and the diameter of the drain openings is less than 20% of the diameter of the input opening, thereby inhibiting passage of intact cells into the drain channels. 
     
     
         62 . The chamber of  claim 53 , wherein the drain channels contain beads or a filter, thereby inhibiting passage of intact cells into the drain channels. 
     
     
         63 .- 64 . (canceled) 
     
     
         65 . The chamber of  claim 53 , at least 50 microns across at its narrowest diameter. 
     
     
         66 . (canceled) 
     
     
         67 . The chamber of  claim 53 , comprising a convex lower surface. 
     
     
         68 . The chamber of  claim 53 , comprising a lower surface coated with a substance that promotes cell adhesion. 
     
     
         69 . The chamber of  claim 68 , wherein the substance is an extracellular matrix. 
     
     
         70 . The chamber of  claim 68 , wherein the substance is fibronectin. 
     
     
         71 . The chamber of  claim 53 , wherein the chamber, the input channel, and the drain channels are filled with fluid. 
     
     
         72 . The chamber of  claim 53 , containing at least two eukaryotic cells. 
     
     
         73 . A plurality of four or more chambers according to  claim 53 , each having its own separate input channel, wherein each of the input channels is configured to supply cells from a common source. 
     
     
         74 .- 76 . (canceled) 
     
     
         77 . A method of maintaining cells in culture, comprising delivering cells to a chamber according to  claim 53 , supplying nutrient medium into the chamber through the input channel or a separate reagent channel, and supplying gas to the gas permeable membrane through the supply channel. 
     
     
         78 . The method of  claim 77 , further comprising individually selecting the cells delivered to the chamber from a cell mixture. 
     
     
         79 . A method of extracting intracellular components from one or more cells that are present in a chamber according to  claim 53 , comprising lysing the cells by delivering a fluid that causes cell lysis into the chamber, and retrieving products of the lysis from the chamber. 
     
     
         80 . The method of  claim 79 , wherein the products are retrieved through the drain channels. 
     
     
         81 . The method of  claim 79 , wherein the products comprise nucleic acid. 
     
     
         82 . The method of  claim 79 , further comprising subjecting the products to a chemical or biochemical reaction. 
     
     
         83 .- 124 . (canceled)