User-Friendly Approach to Gain Isolation of Interphase Cells for Fluorescence in Situ Hybridization

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Background:Fluorescence in situ hybridization (FISH) can be performed on metaphase- and/or interphase cells. Metaphase cells can exclusively be obtained and studied after time-consuming cell culture. Therefore, this technique is suitable only for cells derived from peripheral blood, bone marrow or fibroblasts, the latter including skin, tumour or "prenatal tissues", such as the amnion or chorion. On the other hand, interphase cells can be gained from many different cells and even without cell culture. Methods:Here, we report an approach by which interphase cells can be easily obtained from the skin surface, simply and without causing any pain or injury to the patient. This straightforward procedure involves adding a few microliters of fluid to the patient's skin (e.g. the forearm) and subsequently, using the edge of a slide to collect skin cells together with the fluid by carefully scratching along the wetted skin region. The collected fluid is distributed across the slide surface and air-dried. Results:Applying this simple cell collection approach, several hundred skin cells are obtained. The result can be evaluated by phase contrast microscopy and regions with the highest density of cells can be selected for interphase-FISH. Here, we provide proof-of-principle of this technique. Also, the cells obtained using the described method can be used for DNA-based molecular genetic tests. Conclusions: A new and straightforward method for acquisition of interphase cells is presented. It can be used to complement the available approaches for the analysis of different tissues in cases of questionable low-level mosaics detected in peripheral blood testing.

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