Preservation of Slides

[Rat-Terrier]

What is the best way to produce long-lasting cytological slides. I have been using Dif-Quik to stain and have been viewing with oil. After a couple of days, the cells start looking less like themselves.

I remember in school we had slides with cover slips permanently fixed onto the slide. These were great because you could clean the oil off of them after viewing without damaging the sample. Is there a way to permanently mount a cover slip onto slides so that cleaning would work better? Would this help with the cell degeneration?

Thanks in advance!
-RT

[Peter]

Hi Rat-Terrier,
What cells (tissues) are you viewing? (Presumably Live).
Generally to make a permanent slide the cells need to be fixed (with a killing/fixing solution), stained, dehydrated, possibly/probably cleared (dehydrating agent removed), then mounted in a resinous or similar medium.
If you tell us more of what you are working with we may be able to give more specific and detailed information.
Hope this helps.
Peter.

[Rat-Terrier]

Peter,

Thanks for the reply! I use a quick fix before staining. The cells are from various soft tissues and from blood. For dehydration, would air drying work or do you need to use alcohol or something of that nature? Can dehydration damage cells, and if so, what dehydration method would cause the least damage? Are there any commonly available materials that you would suggest for mounting?

Thanks again!
-RT

[mrsonchus]

Hi RT, if you're using smears then air-drying will be fine for dehydration, if not, it's an alcohol (IPA) series for the tissue/s. You really need to give more details to enable a useful answer to be given - how are you cutting your tissues, are you using the paraffin method for example or are your samples thick? Smears for blood? How are you attaching the tissues to the slides? These are all very relevant details if an outline for the tissue-processing is required...
In isolation the question is hard to answer in a useful way I'm afraid. The wrong advice could lead you to carry out a lot of work for no reward - presumably the 'soft tissues' are zoological? This fact for example has a great bearing on the chemicals used, fixing for example is quite different for animal and plant material...

A broad outline of what you are attempting to do would surely enable folk to give you some very good advice on this forum, there are a lot of very knowledgeable and experienced folk here.

Good luck.

[Peter]

Hi Rat-Terrier,
Blood smears are traditionally air dried then stained with Wright's stain (which contains enough methyl alcohol to fix the smear), then after rinsing are air dried again before mounting (permanently). Other high protein smears may be treated similarly.
A squash is much more difficult to make permanent as removing the coverslip (to fix, dehydrate, etc) will disrupt the carefully prepared squash; some success may be achieved by freezing the slide then prising off the coverslip before processing.
Again a more detailed description of what you are doing will help us help you.
Hope this helps.
Peter.