title: Optimizing optical flow cytometry for cell volume-based sorting and analysis.
abstract: Cell size is a defining characteristic central to cell function and ultimately to tissue architecture. The ability to sort cell subpopulations of different sizes would facilitate investigation at genomic and proteomic levels of mechanisms by which cells attain and maintain their size. Currently available cell sorters, however, cannot directly measure cell volume electronically, and it would therefore be desirable to know which of the optical measurements that can be made in such instruments provide the best estimate of volume. We investigated several different light scattering and fluorescence measurements in several different cell lines, sorting cell fractions from the high and low end of distributions, and measuring volume electronically to determine which sorting strategy yielded the best separated volume distributions. Since we found that different optical measurements were optimal for different cell lines, we suggest that following this procedure will enable other investigators to optimize their own cell sorters for volume-based separation of the cell types with which they work.
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, United States of America.
authors.forename: Amit
authors.initials: A
authors.lastname: Tzur
authors.forename: Jodene K
authors.initials: JK
authors.lastname: Moore
authors.forename: Paul
authors.initials: P
authors.lastname: Jorgensen
authors.forename: Howard M
authors.initials: HM
authors.lastname: Shapiro
authors.forename: Marc W
authors.initials: MW
authors.lastname: Kirschner
mesh_terms: D000818:Animals; D002460:Cell Line; D002469:Cell Separation / Q000379:methods*; D048429:Cell Size; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D012542:Scattering, Radiation