Hello everybody
I use to cryosection pieces of human skin after PFA 4% fixation and sucrose 30% incubation.
pieces arethen embedded in cryomatrix then kept frozen at -80°.
Then sections of 10 µm are cut with a cryostat and sections are collected on superfrost plus slides, let for one hour to dry and stored at -80°.
Since abt 1 year I have troubles with this : when I rehydrate the slides with PBS before immunofluo laballing, the sections begin to float instead of staying stuck to the slide....
I used to realize this technique since a long time w/o any trouble and I really don't see what could explain this and what to do to avoid this floating of sections...?
thanks for your advices
Sounds like your slides are in need of coating with albumin fixative or another tissue adhesive. Tissue sections do not strongly adhere to glass. If your slides are clean, but uncoated you will continue to have this problem.
Hello!
I had the same problem with brain tissue sections. I changed to polysine microscope slides from VWR and significantly reduced this occurence.
Best regards and good luck!
Dear Corre,
Check the expiration date on your lot of slides. Unfortunately the charge lasts only about a year, so it may be as simple as buying new slides if
Superfrost plus always worked for you. If you have irreplaceable slides that you are trying to rescue (I have), you can try a few things. First, make sure you bring the slides to room temperature in a sealed container so that you don’t get cryocondensation on them. In my experience, the water will cause the sections detach. l have also found it helpful to keep the slides as flat as possible, and to remove solutions as gently as possible. You may also try adding a fixation step to refix the section to the slide, before you start your staining protocol. I use 4% paraformaldehyde, usually in a phosphate buffer, but whatever works for your protocol, and rinse well.
Good Luck! Jill
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