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Annals of Clinical & Laboratory Science 37:85-88 (2007)
© 2007 Association of Clinical Scientists

Technical Note

Fluorescence In Situ Hybridization (FISH) Using Snap Frozen Buffy Coat

Hun Jung Min1,4,5, Jinkyoung Lee2, Jung Eun Choi1, Su Shin3 and Dong Soon Lee1,4,5
1 Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea2 Department of Laboratory Medicine, Korea Cancer Center Hospital Seoul, Korea3 Department of Laboratory Medicine, Boramae Hospital, Seoul, Korea4 Cancer Reseach Institute, Seoul National University College of Medicine, Seoul, Korea5 National Research Laboratory for Molecular Cell Imaging, Seoul, Korea

Address correspondence to Dong Soon Lee, M.D., Department of Laboratory Medicine, Seoul National University College of Medicine, 28 Yongon-dong, Chongno-gu, Seoul, 110-744, Korea; tel 82 2 2072 3986; fax 82 2 745 6653; e-mail: soonlee{at}plaza.snu.ac.kr or leukemia{at}plaza.snu.ac.kr.

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°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.

Keywords: cytogenetics, chromosome probes, buffy coat, FISH analysis






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