METHOD FOR DETECTING RARE EVENT
Provided are methods, compositions and kits for efficiently and accurately identifying rare events in a biological sample.
1 . A method for identifying proliferative disorders in a biological sample, comprising:
contacting an obtained source of cells with a binding agent specific for a cell specific marker associated with a proliferative disorder and expressed by at least some of the cells, wherein the binding agent is bound to a magnetic bead and wherein the binding agent binds to cells in the source expressing the cell specific marker;
separating cells bound by the binding agent from the source thereby obtaining a sub-population of cells enriched for the cell specific marker associated with the proliferative disorder;
placing the enriched sample on a microscope slide;
automatically scanning the microscope slide at a plurality of coordinates using a microscope;
automatically obtaining a plurality of images at locations on the microscope slide that comprise the enriched sample; and
processing each of the plurality of images to identify the proliferative disorder, including transforming the images to a different color space, thresholding the images, and determining the presence of one or more regions of connected pixels having the same color in the thresholded images.
2 . The method according to claim 1 , wherein the binding agent is an antibody.
3 . The method according to claim 1 , wherein the sub-population is enriched for carcinoma cells.
4 . The method of claim 1 , wherein the separating is done by positive selection.
5 . The method of claim 1 , wherein the separating is done by negative selection.
6 . The method of claim 2 , wherein the antibody is monoclonal or polyclonal.
7 . The method of claim 2 , wherein the antibody recognizes an epithelial marker.
8 . The method of claim 2 , wherein the antibody is selected to avoid cross reactivity with the beads.
9 . The method of claim 3 , wherein the carcinoma cells are from peripheral blood.
10 . The method of claim 1 , further comprising:
(a) automatically identifying a coordinate of the proliferative disorder; and
(b) automatically acquiring an image of the proliferative disorder, at the location coordinates.
11 . The method of claim 1 , wherein the proliferative disorder is detected by immunohistochemistry.
12 . The method of claim 1 , wherein the proliferative disorder is detected by in situ hybridization.
13 . The method of claim 1 , wherein the cell specific marker is detected by immunohistochemistry, in situ hybridization, staining or a combination thereof.
14 . The method of claim 1 , wherein the image is a digital image.