Rapid Clonal Growth Measurements at the Single-Cell Level: Gel Micrdorplets and Flow Cytometry

Biotechnology, September 1991, vol. 9, pp. 873-876

James C. Weaver, Jonathan G. Bliss, Kevin T. Powell, Gail I. Harrison and Gregory B. Williams

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 µm 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 sub-population 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.

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