Fluorescence in situ hybridization (FISH) using snap frozen buffy coat.
We report the successful application of interphase fluorescent in situ hybridization (FISH) using extracted nuclei from snap frozen buffy coat stored for the purpose of molecular analysis. Included in this study were 30 frozen buffy coat specimens, ranging from <1 day to 3 yr old. Thawing of the buffy coat at 37 degrees C was followed by lysing erythrocytes with lysing solution (LYSE S III DIFF, Coulter Corp.). Cells were fixed and slides were prepared according to manufacturer's FISH protocol. All specimens from the 30 frozen-thawed buffy coats were suitable for FISH evaluation, and signal intensities were not significantly related to the age of specimens. In conclusion, interphase FISH is feasible using frozen-thawed buffy coat; the technique will be useful for retrospective molecular cytogenetic analysis of hematologic malignancies.
Min
Hun Jung
HJ
Department of Laboratory Medicine, Seoul National University College of Medicine, and Korea Cancer Center Hospital Seoul, 110-744, Korea.
Lee
Jinkyoung
J
Choi
Jung Eun
JE
Shin
Su
S
Lee
Dong Soon
DS
D001773:Blood Cells / Q000166:cytology*; D001793:Blood Preservation / Q000379:methods*; D002467:Cell Nucleus / Q000235:genetics*; D015925:Cryopreservation / Q000379:methods; D005069:Evaluation Studies as Topic; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007399:Interphase / Q000235:genetics