A simple electronic volume cell sorter for clonogenicity assays.
A single-parameter electronic volume flow cell sorter that can be easily and inexpensively constructed using existing technology is described. The instrument is designed for ease and flexibility of operation, including such features as a large open area for recovering sorted cells into a variety of dishes or vessels; a remote, electrically activated fluidics system; a mechanism for heating or cooling samples during sorting; a simple arrangement for monitoring and adjusting the sorting control parameters; and an interface to a standard IBM personal computer for data acquisition, analysis, and control of the sorting windows. Several researchers in our laboratory now routinely use this sorter for plating precise numbers of cells directly into culture dishes in an aseptic manner for clonogenicity assays. The instrument can sort cells at rates of up to approximately 2,000 per second with greater than 80% sorting efficiency and no cytotoxicity. An advantage of this system is that the sorting windows can be set to exclude acellular debris and include either the entire cell volume distribution or a subset thereof. Applications of the instrument are detailed, including 1) precise cell plating for low-dose survival studies, 2) separation of cells into age compartments, and 3) rapid inoculation of single cells into multiwell dishes for cloning studies. Advantages of this technology for cell survival studies are detailed, along with some limitations to its applicability.
Freyer
J P
JP
Cell Biology Group, Los Alamos National Laboratory, New Mexico 87545.
Wilder
M E
ME
Schor
P L
PL
Coulter
J
J
Raju
M R
MR
D000818:Animals; D002469:Cell Separation; D004273:DNA, Neoplasm / Q000032:analysis*; D001330:Electronic Data Processing; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D014407:Tumor Cells, Cultured / Q000032:analysis / Q000166:cytology*
D004273:DNA, Neoplasm