Label-free cell segmentation using phase contrast and brightfield imaging

Abstract

The disclosure provides example methods that include a processor: (a) generating at least one phase contrast image of a biological specimen comprising one or more cells centered around a focal plane for the biological specimen; (b) generating a confluence mask in the form of a binary image based on the at least one phase contrast image; (c) receiving a first brightfield image of the biological specimen at a defocusing distance above the focal plane and a second brightfield image of the biological specimen at the defocusing distance below the focal plane; (d) generating a cell image of the biological specimen based on the first and second brightfield image; (e) generating a seed mask based on the cell image and the phase contrast image; and (f) generating an image of the biological specimen showing a cell-by-cell segmentation mask based on the seed mask and the confluence mask.