单细胞水平的快速克隆生长测量：凝胶微滴与流式细胞术。

Rapid clonal growth measurements at the single-cell level: gel microdroplets and flow cytometry.

作者信息

Weaver J C, Bliss J G, Powell K T, Harrison G I, Williams G B

机构信息

Harvard-M.I.T. Division of Health Sciences and Technology, Massachusetts, Cambridge 02139.

出版信息

Biotechnology (N Y). 1991 Sep;9(9):873-7. doi: 10.1038/nbt0991-873.

DOI:10.1038/nbt0991-873

PMID:1367361

Abstract

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.

摘要

我们描述了一种新的通用方法，用于快速测量细胞群体中大量单个成员的克隆生长。该方法基于在凝胶微滴（GMD；此处为琼脂糖；直径20至90微米）中对单个集落形成单位进行微培养，凝胶微滴足够坚固，可像细胞一样进行操作，并且对感兴趣的分子具有扩散透明性。流式细胞术可对包埋在GMD中的微菌落进行快速测量，并允许进行亚群分析。本文用哺乳动物、真菌和细菌物种演示了该方法。其他结果表明可快速确定混合物种样品中的耐药亚群，以及小鼠杂交瘤的营养敏感性。