title: Cell sorting in a Petri dish controlled by computer vision.
abstract: 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.
authors: Institute of Experimental Medicine of the Hungarian Academy of Sciences, Budapest, Hungary.
authors: Z
authors: Z
authors: Környei
authors: S
authors: S
authors: Beke
authors: T
authors: T
authors: Mihálffy
authors: M
authors: M
authors: Jelitai
authors: K J
authors: KJ
authors: Kovács
authors: Z
authors: Z
authors: Szabó
authors: B
authors: B
authors: Szabó
mesh_terms: D016475:3T3 Cells; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology; D002460:Cell Line; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D015603:Keratinocytes / Q000166:cytology; D051379:Mice; D008822:Mice, Transgenic; D017628:Microglia / Q000166:cytology; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D018715:Microscopy, Video; D015203:Reproducibility of Results
chemical_list: D049452:Green Fluorescent Proteins