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Published on: March 25, 2009
Fluorescence in situ hybridization (FISH) using snap frozen buffy coat
Hun Jung Min1, Jinkyoung Lee, Jung Eun Choi
1Department of Laboratory Medicine, Seoul National University College of Medicine, and Korea Cancer Center Hospital Seoul, 110-744, Korea.
Annals of Clinical and Laboratory Science
|February 22, 2007
Summary
Interphase fluorescent in situ hybridization (FISH) is feasible using frozen-thawed buffy coats. This technique enables retrospective molecular cytogenetic analysis of hematologic malignancies, even with older specimens.
Area of Science:
- Molecular Biology
- Cytogenetics
- Hematology
Background:
- Storing buffy coat specimens is crucial for molecular analysis.
- Interphase fluorescent in situ hybridization (FISH) is a key cytogenetic technique.
- Evaluating the utility of frozen buffy coat for FISH is important for retrospective studies.
Purpose of the Study:
- To assess the feasibility of interphase FISH using frozen-thawed buffy coat specimens.
- To determine if specimen age affects FISH signal intensity.
- To establish the utility of this method for molecular cytogenetic analysis.
Main Methods:
- Utilized 30 frozen buffy coat specimens aged up to 3 years.
- Extracted nuclei from snap-frozen buffy coats.
- Applied interphase FISH protocols after thawing and erythrocyte lysis.
Main Results:
- All 30 frozen-thawed buffy coat specimens were suitable for FISH evaluation.
- Specimen age did not significantly impact FISH signal intensity.
- Successful application of interphase FISH on frozen-thawed buffy coat.
Conclusions:
- Interphase FISH is a viable technique for frozen-thawed buffy coat samples.
- This method supports retrospective molecular cytogenetic analysis of hematologic malignancies.
- The age of frozen buffy coat specimens does not hinder FISH analysis.
