Fluorescence-activated cell sorting (FACS) applying flow
cytometry to separate cells on a molecular basis is a widespread
method. We demonstrate that both fluorescent and unlabeled live
cells in a Petri dish observed with a microscope can be
automatically recognized by computer vision and picked up by a
computer-controlled micropipette. This method can be routinely
applied as a FACS down to the single cell level with a very
high selectivity. Sorting resolution, i.e., the minimum distance
between two cells from which one could be selectively removed
was 50-70 micrometers. Survival rate with a low number of 3T3
mouse fibroblasts and NE-4C neuroectodermal mouse stem cells was
66 +/- 12% and 88 +/- 16%, respectively. Purity of sorted
cultures and rate of survival using NE-4C/NE-GFP-4C co-cultures
were 95 +/- 2% and 62 +/- 7%, respectively. Hydrodynamic
simulations confirmed the experimental sorting efficiency and a
cell damage risk similar to that of normal FACS
