Background: It is widely known that the efficiency of fluorescence in situ hybridization (FISH) probes applied to formalin-fixed, paraffin-embedded tissues is affected by the conditions under which the tissues are fixed and embedded. However, relatively few studies address exactly how tissue archiving conditions affect the performance of FISH probes. We report our experience based on use of an ALK FISH probe, during the validation of its diagnostic utility.
Methods: We applied the probe to 77 formalin-fixed, paraffin-embedded tissue blocks archived from 1991 through to 2000, and studied the interrelationship between the archival age (which ranged up to 10 years), type and condition of tissue, duration required for optimum hydrolysis, and obtainability of hybridization signals.
Results: We found that as archival age and tissue collagen content increased, not only did hydrolysis times have to be prolonged in order to yield interpretable hybridization signals, but also the likelihood of blocks becoming non-signaling increased. The most striking positive correlations were seen between the archival age of signaling lymphoid blocks and their requisite hydrolysis times.
Conclusions: The difficulty in applying FISH on archival tissue increases with its archival age and collagen content, and may necessitate changes in laboratory protocol accordingly.
Anaplastic Lymphoma Kinase Anaplastic Large Cell Lymphoma Hydrolysis Time Signaling Block Archival Tissue
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Funding for this study came from the Health Services Development Program, Ministry of Health, Singapore.
We would like to thank Stella Chee, BSc, of the Molecular Diagnosis Centre, Department of Laboratory Medicine, National University Hospital (NUH), for her assistance in performing the FISH assays, as well as Matsuni bin Hamzah of the Special Histology Section, Department of Pathology, National University of Singapore, and Wendy Ang of the Department of Pathology, NUH, who helped in configuring the graphs and tables, respectively.
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