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ORIGINAL RESEARCH article
Front. Genet.
Sec. Molecular Cytogenetics
Volume 16 - 2025 | doi: 10.3389/fgene.2025.1569308
Hapten labeled DNA-probes can be stored and used in fluorescence in situ hybridization for decades
Provisionally accepted- 1Friedrich Schiller University Jena, Jena, Germany
- 2University Hospital Jena, Jena, Thuringia, Germany
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In molecular cytogenetics, fluorescence in situ hybridization (FISH) is the main technique used. In both research and diagnostics, FISH depends on DNA probes that are well defined and mapped and that lead to brilliant signals with little background visible in metaphases and/or interphases. Such probes are either ready for use and commercially available or are provided as unlabeled DNA. The latter can be obtained by flow sorting, microdissection or by cloning DNA segments into appropriate bacterial vectors. Labeling can be done with nonfluorescent or fluorescent haptens. According to international guidelines, such FISH probes must have a minimum shelf life which is only between 2 and 3 years in human genetic diagnostics. The Molecular Cytogenetics Laboratory reporting here has been purchasing, producing, using and storing FISH probes since the 1990s. For this study, the available stock of approximately 25,000 labeled probes was screened. 581 FISH probes labeled and approved 1 to 30 years before another use were selected for this study; 75 of these were commercially available probes labeled 1 to 20 years ago. All of these probes, stored in the dark at -20°C, worked perfectly well in the FISH method. While only slight to no differences in exposure times were observed over the years for self-labeled, homemade probes, commercial probes labeled with SpectrumOrange had shorter exposure times and maintained them over the years. DNA probes labeled with SpectrumAqua / diethylaminocoumarin showed some bright labeling for the first three years and then faded. Accordingly, it can be assumed that self-labeled, homemade and commercial FISH probes can be stored stably in the dark and at -20°C for at least 30 years or longer. There is no need to test approved probes on a slide after the official expiry date. In practice, this means that a FISH probe tube that has been approved can even be used in diagnostics until it is empty; there is no need to dispose of the expensive probes at any point due to their age.
Keywords: fluorescence in situ hybridization (FISH), molecular cytogenetics, Indirect labeling, Direct labeling, commercial probes, homemade probes
Received: 31 Jan 2025; Accepted: 18 Apr 2025.
Copyright: © 2025 Liehr, Padutsch and Kankel. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Thomas Liehr, Friedrich Schiller University Jena, Jena, Germany
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