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- Title
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.
- Authors
Sederberg, Maria C.; Guangyu Gu; Sarah T. South
- Abstract
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 pf probe mixture required per area of hybridization was decreased from 10 μl to 3 ~μl. 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 of the 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.
- Subjects
BIOLOGICAL reagents; FLUORESCENCE in situ hybridization; METHANOL; CHEMICAL synthesis; FATTY acids; PHYSIOLOGICAL effects of acetic acid
- Publication
Journal of the Association of Genetic Technologists, 2013, Vol 39, Issue 1, p7
- ISSN
1523-7834
- Publication type
Article