Methods of using non-rare cells to detect rare cells

Abstract

The invention provides seminal computational approaches utilizing data from non-rare cells to detect rare cells, such as circulating tumor cells (CTCs). The invention is applicable at two distinct stages of CTC detection; the first being to make decisions about data collection parameters and the second being to make decisions during data reduction and analysis. Additionally, the invention utilizes both one and multi-dimensional parameterized data in a decision making process.

1 - 53 . (canceled). 
     
     
         54 . A method for detecting a circulating tumor cell (CTC) in a sample from a subject comprising:
 a) plating a non-enriched sample comprising nucleated blood cells onto a slide, said non-enriched sample suspected of comprising circulating tumor cell (CTCs) and non-CTCs wherein said non-CTCs are present at a concentration that is at least 10 times that of the CTCs;   b) contacting the sample with a first immunuofluorescently detectable agent targeting a positive marker and a second immunuofluorescently detectable agent targeting a negative marker;   c) scanning the slide with fluorescent scanning microscopy to generate a cell image of the nucleated cells in the sample; and   d) identifying the CTC in the sample based on based on assessment of said detectable agents and morphology in said cell image.   
     
     
         55 . The method of  claim 54 , wherein said positive marker targets a specific cell type or tissue. 
     
     
         56 . The method of  claim 55 , wherein said positive marker targets a specific cell type. 
     
     
         57 . The method of  claim 56 , wherein said specific cell type is epithelial. 
     
     
         58 . The method of  claim 57 , wherein said positive marker is cytokeratin. 
     
     
         59 . The method of  claim 54 , wherein said negative marker targets non-CTCs. 
     
     
         60 . The method of  claim 59 , wherein said non-CTCs are white blood cells. 
     
     
         61 . The method of  claim 60 , wherein said negative marker is CD45. 
     
     
         62 . The method of  claim 54 , further comprising a third immunofluorescently detectable agent. 
     
     
         63 . The method of  claim 62 , wherein said third immunofluorescently detectable agent targets a content marker. 
     
     
         64 . The method of  claim 63 , wherein said content marker is androgen receptor (AR).