Multiparameter FACS assays to detect alterations in cellular parameters and to screen small molecule libraries

Abstract

The invention relates to novel methods of detecting alterations in cell cycle regulation in a cell or a cell population and screening for agents capable of modulating cell cycle regulation through the use of multiparameter assays and a fluorescence-activated cell sorter (FACS) machine.

1 - 10 . (canceled)  
     
     
         11 . A method comprising: 
 a) contacting a population of cells with a candidate bioactive agent to produce contacted cells; and    b) sorting said contacted cells on the basis of at least five different cellular parameters using a fluorescence-activated cell sorter (FACS).    
     
     
         12 . The method of  claim 11 , wherein said at least five cellular parameters includes detection of at least one of: forward light scattering, side light scattering, light refraction, fluorescent dye release, fluorescent dye uptake, and a fluorescent protein.  
     
     
         13 . The method of  claim 11 , further comprising identifying a cell having a phenotype indicated by said at least five cellular parameters.  
     
     
         14 . The method of  claim 13 , wherein said phenotype is a cell cycle phenotype.  
     
     
         15 . The method of  claim 11 , wherein said method further comprises subjecting said cells to a phenotype-inducing stimulus prior to said sorting step b).  
     
     
         16 . The method of  claim 11 , wherein said candidate bioactive agent is a nucleic acid comprising a sequence that encodes a polypeptide.