Optimized fixation and storage conditions for FISH analysis of single-cell suspensions.
J Histochem Cytochem
; 46(8): 971-4, 1998 Aug.
Article
en En
| MEDLINE
| ID: mdl-9671447
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. (J Histochem Cytochem 46971-973, 1998)
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Banco de datos:
MEDLINE
Asunto principal:
Manejo de Especímenes
/
Leucocitos Mononucleares
/
Hibridación Fluorescente in Situ
Tipo de estudio:
Prognostic_studies
Límite:
Humans
Idioma:
En
Revista:
J Histochem Cytochem
Asunto de la revista:
HISTOCITOQUIMICA
Año:
1998
Tipo del documento:
Article
País de afiliación:
Estados Unidos