Rapid clonal growth measurements at the single-cell level:
gel microdroplets and flow cytometry.
Weaver JC; Bliss JG; Powell KT; Harrison GI; Williams GB Biotechnology (N Y) (AL1), 1991 Sep; 9 (9): 873-7
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.