Abstract
The fluorescent vital dye, Hoechst 33342, was used to stain cultured cells prior to assay of antibody dependent complement mediated cytotoxicity. The fluorescence of nonviable dye stained cells is quenched by cellular uptake of trypan blue, but trypan blue excluding cells remain intensely fluorescent. Detection by fluorescence microscopy of one viable prestained cell per 105 unstained cells was accurate and reliable. The technique was found to have sensitivity equal to a clonogenic assay for measuring cytotoxicity. The dye stained cell assay may be used to measure depletion of a selected cell type, when those cells are stained prior to mixing with another cell population. This technique may prove useful to study model systems for depletion of tumor cells or T-cells from bone marrow.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 39-46 |
| Number of pages | 8 |
| Journal | Journal of Immunological Methods |
| Volume | 82 |
| Issue number | 1 |
| DOIs | |
| State | Published - Sep 3 1985 |
Funding
The ability of cultured cell lines to form colonies when suspended in soft agar has also been used as a functional assay of cell survival. Great sensitivity can be obtained with the clonogenic assay technique. However, results are not immediately * Supported in part by grants from the United Cancer Council of Greater Rochester, Inc. and Grant RR-05403 from the United States Public Health Service. C.N.F. is a recipient of an American Cancer Society Junior Faculty Research Award. 1 To whom correspondence should be addressed.
Keywords
- Hoechst 33342
- clonogenic assay
- complement mediated cytotoxicity
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology