FISHing Efficiency: A Streamlined Approach to Reducing the Time and Reagents Required to Perform Fluorescence in situ Hybridization Assays on Methanol: Acetic Acid-fixed, Cultured Cells

摘要

Recent assessments run in our laboratory indicate that some steps in a standard fluorescence in situ hybridization (FISH) assay on methanol: acetic acid-fixed cultured cells can be significantly reduced or even eliminated while still producing high-quality signals that are concordant with results obtained from traditional FISH assay methods.The streamlined protocol increased the number of probes per slide from two to three. The requirement for a pretreatment was removed and the amount of probe mixture required per area of hybridization was decreased from 10 ul to 3 ul.Probes from five FISH panels, using a total of nineteen probes, were selected. The panels included acute myeloid leukemia (AML), eosinophilia (EOS), multiple myeloma (MM), myelodysplastic syndrome (MDS), and chronic lymphocytic leukemia (CLL), plus BCR/ABL1.Scores and hybridization quality for the streamlined protocol were compared against the scores and hybridization quality observed on the traditional slide run for clinical assessment. The streamlined assay worked well on all probes, with the exception ofthe probes involved in the chronic lymphocytic leukemia (CLL) panel.However, it was observed that reducing the amount of probe mixture with the CLL panel did not affect the quality of the signal, which reduced the cost of probe per sample if not the overall amount of time.