Hi everyone, In the diagnostic workflow of the test, we often need to reuse cytology slides, which requires removing the coverslip previously mounted over the sample. Currently, we use xylene, and the slides are left immersed for up to 72 hours until the coverslip detaches. However, we are looking for a faster and more efficient method to optimize our routine. Has anyone had experience with alternative solvents or techniques that are more effective for removing the coverslip and the mounting medium (typically resin or Canada balsam)? Any suggestions based on practical experience or literature references would be greatly appreciated.
Hello Miriana,
obviously the problem is that any solvent can only penetrate at the narrow edge between the slide and the cover glass. To speed this up, we scratch the cover glass crosswise with a diamond pin. Of course, this damages the specimen, but this does not play a role in our field of activity, as we no longer have to microscopy the histological material afterwards and only use it for DNA analysis. Maybe you proceed differently, then this advice would be useless for you.
With kind regards from Mainz, Germany
Klaus
Klaus Berkefeld Thank you very much, Klaus, for your reply and for sharing your approach! Your point about solvent penetration being limited to the narrow edge between the slide and coverslip makes perfect sense — and your technique of scratching the coverslip with a diamond pin is very clever to accelerate the process. In our case, we still need to preserve the cytological material for further molecular analysis (RNA and DNA extraction for molecular analises), but your comment made me reflect that scratching the coverslip probably wouldn’t compromise the biological material, as we don’t rely on the microscopic morphology afterward. May I ask: how long do you typically leave the slides immersed in xylene before the coverslip detaches? And do you keep the solvent at room temperature or use mild heating to speed up the process? Thanks again for your valuable input!
Hello Miriane,
since we obtain our specimens from various sources and sometimes don't know exactly which embedding medium was used, I can't provide general information. However, it's usually sufficient to leave the specimens covered with xylene in a Petri dish overnight after scoring, allowing the coverslip fragments to be carefully removed. To avoid additional problems with solvent vapors, we don't heat this setup.
As you can see, we only do this occasionally, and I don't know if this procedure would be suitable for high-throughput applications.
With kind regards
Klaus
Klaus Berkefeld Thank you so much for the additional details, Klaus — that’s very helpful! Indeed, your point about the variability of mounting media is important, and I appreciate your clarification about not using heat and letting the slides sit overnight. We’ll definitely consider testing this approach with a few slides, including scoring the coverslip beforehand, and see how it performs in our context. Thanks again for your valuable insights and your time!
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