Patent ID: 8175371

Claim:
A method for classifying a specific cell as one of the following four types of cells, a viable cell, a necrotic cell, an early apoptotic cell in which a cellular membrane of the cell is still intact, and a late apoptotic cell in which the cellular membrane of the cell is not intact, using only a single nuclear marker and image data from the cell, the method comprising the steps of: (a) exposing the specific cell to only a single nuclear marker that will bind to DNA in a nucleus of the specific cell in the event that the cellular membrane of the specific cell is not intact; (b) collecting image data from the specific cell, the image data comprising: (i) a darkfield image of the specific cell, the darkfield image corresponding to a side scatter image of the specific cell; (ii) a brightfield image of the specific cell; and (iii) a fluorescent image of the specific cell, the fluorescent image enabling the location of the single nuclear marker in the specific cell to be determined; (c) using the brightfield image of the specific cell and the fluorescent image of the specific cell to calculate a cytoplasmic size of the specific cell; (d) analyzing the fluorescent image of the specific cell to determine if the nuclear marker is present in the nucleus of the specific cell; (e) using the darkfield image of the specific cell to calculate a spatial scatter frequency metric indicative of a granularity of the specific cell; and (f) classifying the specific cell as one of the four types of cells based on the following: (i) the cytoplasmic size of the specific cell; (ii) the darkfield scatter frequency metric of the specific cell; and (iii) the presence of the nuclear marker in the nucleus of the specific cell.