[The detection of apoptosis cells by TUNEL stain in peripheral blood smears].
We have determined the fixative conditions for TUNEL stain in peripheral blood smears. Murine IL-3 dependent B cell line (BAF-3) was used as positive control. The intensity of fixing solutions effected on the positivity/negativity for TUNEL stain. Alive cells showed positivity for TUNEL stain (false positive), when 70% methyl alcohol, acetone or carnoy fluid was used. In contrast to the strong fixation by high concentrated glutaraldehyde and the phosphate buffer, denatured cells showed to be negative (false negative). The fixation by 0.4% glutaraldehyde and 70% methyl alcohol for 15 min at room temperature, followed by the treatment by 20 micrograms/ml proteinase K for 15 min at 37 degrees C was most suitable for TUNEL stain to detect the apoptotic cells in peripheral blood smears. The frequency of TUNEL positive cells increased significantly in peripheral blood cells treated by apoptosis induced reagents, or infected by CMV. This is a new approach for the techniques to detect the apoptotic cells in peripheral blood smears.
Okano
K
K
School of Allied Health Sciences, Yamaguchi University, Ube.
Yamamoto
K
K
Ueda
J
J
Yamashiro
Y
Y
Araki
M
M
Ishida
Y
Y
D017209:Apoptosis; D001773:Blood Cells / Q000166:cytology*; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis; D004586:Electrophoresis; D006801:Humans; D020287:In Situ Nick-End Labeling
D004247:DNA