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

Sarah Murrell-Bussell, Dianne Nguyen, Wendy D. Schober, Jeffrey Scott, Joe Leigh Simpson, Sherman Elias, Farideh Z. Bischoff, Dorothy E. Lewis

Research output: Contribution to journalArticle

7 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)971-973
Number of pages3
JournalJournal of Histochemistry and Cytochemistry
Volume46
Issue number8
DOIs
StatePublished - Jan 1 1998

Keywords

  • Cell fixation methods
  • Flow cytometry
  • Fluorescence in situ
  • Hybridization (FISH)
  • Paraformaldehyde (PF)

ASJC Scopus subject areas

  • Anatomy
  • Histology

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