I have some bacterial smears that have been Gram stained (crystal violet-lugol-safranin). I would like to preserve those as permanent slides for record keeping, but I am unsure on how to perform this. Should I just add a coverslip and seal the borders or do the slides require some other more specific treatment? Thank you for the help!
Hi Liany,
I assume the section are thoroughly dry prior to placing emersion oil on them for microscopy. Once microscopy is finished place the slides in xylene for a few minutes to remove the immersion oil and use a polystyrene mountant for coverslipping (e.g. DePeX). Once coverslipped the slides can be stored indefinitely and immersion oil can be place directly on the surface of the coverslip for further microscopy.
You can use xylene based Entellan.
after completing the staining you can place your slide into xylene (15 seconds) and then mounting them with entellan and coverslip.
We perform gram in both cytological and histological samples either as first stain or secondary after destaining by hydrochloric acid. For both settings conventional xylene-based mounting media work without fading.
Air dry then clear in xylene and mount in DePeX.
Avoid all alcohols as this will remove the stain.
Thank you all for the replies. I've never done this before.. Is there any chance I could get a detailed protocol online?
Hi Liany,
I assume the section are thoroughly dry prior to placing emersion oil on them for microscopy. Once microscopy is finished place the slides in xylene for a few minutes to remove the immersion oil and use a polystyrene mountant for coverslipping (e.g. DePeX). Once coverslipped the slides can be stored indefinitely and immersion oil can be place directly on the surface of the coverslip for further microscopy.
Hey Jason,
I used a tape to hold a coverslip on a smear today and it allowed me to take a look at the slide without applying immersion oil directly to the sample. In this case, could I skip the xylene step and apply the DePeX at once?
Thanks for your help
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