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Improved technique for fluorescence in situ hybridisation analysis of isolated nuclei from archival, B5 or formalin fixed, paraffin wax embedded tissue

Department of Pathology, Queen's University, Kingston, Ontario, K7L 2V7, Canada

Correspondence to:
Dr K Harrison, Department of Pathology, Richardson Laboratory, Room 201, Kingston, Ontario, K7L 2V7, Canada;
harrison{at}cliff.path.queensu.ca

ABSTRACT
Fluorescence in situ hybridisation (FISH) is an effective method to detect chromosomal alterations in a variety of tissue types, including archived paraffin wax embedded specimens fixed in B5 or formalin. However, precipitating fixatives such as B5 have been known to produce unsatisfactory results in comparison with formalin when used for FISH. This study describes an effective nuclear isolation and FISH procedure for B5 and formalin fixed tissue, optimising the nuclear isolation step and nuclei pretreatments using tonsil and mantle cell lymphoma specimens. The protocol presented can be used to isolate nuclei and perform FISH on B5 or formalin fixed, paraffin wax embedded samples from a variety of tissue types.

Keywords: fluorescence in situ hybridisation; B5; formalin; paraffin wax

Abbreviations: CGH, comparative genomic hybridisation; FISH, fluorescence in situ hybridisation; MCL, mantle cell lymphoma; PBS, phosphate buffered saline; SCC, standard saline citrate

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