Optimized fixation and storage conditions for FISH analysis of single- cell suspensions Article

Murrell-Bussell, S, Nguyen, D, Schober, WD et al. (1998). Optimized fixation and storage conditions for FISH analysis of single- cell suspensions . 46(8), 971-973. 10.1177/002215549804600811

cited authors

  • Murrell-Bussell, S; Nguyen, D; Schober, WD; Scott, J; Simpson, JL; Elias, S; Bischoff, FZ; Lewis, DE

fiu authors

abstract

  • In our protocol to isolate and identify fetal cells in maternal peripheral blood, antibody (Ab)-stained cells are preserved with paraformaldehyde (PF) before batch flow cytometric sorting. However, PF fixation compromises the quality of subsequent interphase fluorescence in situ hybridization (FISH). We therefore examined the effect of PF concentrations and storage time in phosphate-buffered saline (PBS) on the quality of FISH signals. Cells were analyzed for changes in light scatter, morphology, and accessibility of target cell DNA. Fixation in 3% PF for 1 hr was ideal for both flow cytometry and subsequent FISH detection. However, beyond 10 days of storage, FISH quality deteriorated.

publication date

  • January 1, 1998

Digital Object Identifier (DOI)

start page

  • 971

end page

  • 973

volume

  • 46

issue

  • 8