TY - JOUR
T1 - Optimized fixation and storage conditions for FISH analysis of single- cell suspensions
AU - Murrell-Bussell, Sarah
AU - Nguyen, Dianne
AU - Schober, Wendy D.
AU - Scott, Jeffrey
AU - Simpson, Joe Leigh
AU - Elias, Sherman
AU - Bischoff, Farideh Z.
AU - Lewis, Dorothy E.
PY - 1998/1/1
Y1 - 1998/1/1
N2 - 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.
AB - 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.
KW - Cell fixation methods
KW - Flow cytometry
KW - Fluorescence in situ
KW - Hybridization (FISH)
KW - Paraformaldehyde (PF)
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U2 - 10.1177/002215549804600811
DO - 10.1177/002215549804600811
M3 - Article
C2 - 9671447
AN - SCOPUS:0031828223
VL - 46
SP - 971
EP - 973
JO - Journal of Histochemistry and Cytochemistry
JF - Journal of Histochemistry and Cytochemistry
SN - 0022-1554
IS - 8
ER -