METHODS OF USING NON-RARE CELLS TO DETECT RARE CELLS
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).