Rapid clonal growth measurements at the single-cell level: gel microdroplets and flow cytometry.
We describe a new, general method for rapidly measuring clonal growth of large numbers of individual members of a cell population. This method is based on microculture of individual colony-forming units in gel microdrops (GMDs; here agarose; 20 to 90 mu in diameter), which are sufficiently robust to be handled much like cells, and diffusionally transparent for molecules of interest. Flow cytometry provides rapid measurements of GMD-entrapped microcolonies, and permits subpopulation analysis. Here the method is demonstrated with mammalian, fungal and bacterial species. Additional results illustrate rapid determination of a drug-resistant subpopulation in a mixed species sample, and nutrient sensitivity for a murine hybridoma.
Harvard-M.I.T. Division of Health Sciences and Technology, Massachusetts, Cambridge 02139.
J C
JC
Weaver
J G
JG
Bliss
K T
KT
Powell
G I
GI
Harrison
G B
GB
Williams
D000818:Animals; D002175:Candida / Q000166:cytology / Q000254:growth & development*; D002455:Cell Division; D002460:Cell Line; D002999:Clone Cells; D046508:Culture Techniques / Q000379:methods*; D004926:Escherichia coli / Q000166:cytology / Q000187:drug effects / Q000254:growth & development*; D005434:Flow Cytometry / Q000379:methods*; D006825:Hybridomas; D051379:Mice; D008826:Microbial Sensitivity Tests / Q000379:methods; D010400:Penicillin G / Q000494:pharmacology
D010400:Penicillin G