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A method of quantifying cell sorting yield in “real time”
Author(s) -
Osborne Geoffrey W.
Publication year - 2010
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20950
Subject(s) - sorting , biological system , calibration , yield (engineering) , flow cytometry , drop (telecommunication) , biomedical engineering , cytometry , materials science , cell sorting , bead , nanotechnology , computer science , mathematics , biology , microbiology and biotechnology , statistics , engineering , algorithm , composite material , telecommunications
Cell sorting flow cytometers sort cells by applying electrical charges to a stream that forms liquid droplets containing the cells at a set time after sample interrogation. The correct time to apply these charges is determined based on calibration beads and automated technology. The central tenet of this method is that beads accurately indicate the yield of cells sorted using the same instrument parameters. HEK293T cells were incubated with Accudrop™ calibration beads. Cell incorporation of beads by phagocytosis was confirmed by imaging cytometry. Cells containing beads were analyzed and sorted on unmodified commercially available cell sorters that have automated technology for setting drop delay times. Based on cell sorter drop delay times optimized using beads, sorting experiments demonstrate that yield can be assessed in real time using bead loaded cells and existing technology. Here, data presented show that cells have lower sorting yields than indicated using beads. Further, this data show clear trends related to nozzle tip diameter and sorting yield. The present study demonstrates a method to quantify yield “on the fly” during cell sorting, that is separated from drop charge counts and removes the variables associated with yield assessment from collection tubes. These data have implications for the expected recovery of cells from sorting experiments. © 2010 International Society for Advancement of Cytometry