Hy,

you have to warm the glass slides after you plated your tissue slices.

You have to put them on a heat plate and increase the temperature at about 38/39°Celsius for about 15 minutes. then you rinse them in water for histology or in PBS for immunohistochemistry.

Sincerely yours

Stefan

Dear Calum,

I agree with Stefan that some treatment would help after sectioning your tissue. While a little different, I let my slides sit for 1-2 hours at room temperature, immediately after sectioning them onto slides. This has greatly helped retain the tissue sections on the slides and they even hold up during strenuous assays. Hope this is helpful.

-Daniel

You can use a product like Vectabond from Vector labs with the charged slides. Also, really old charged slides are less effective.

Thank you all for your responses! If I were to heat the slides after mounting, would an incubator be sufficient? Or is the idea to only heat the slides from the bottom? I have just purchased new slides so I'm hoping their age isn't affecting them.

Hy Calum,

i used a heat plate for the slides.

But an incubator is also ok.

Usually the date untill you can use the super frost slides is labelled on the package.

Good luck

Stefan

FYI. I have heard that effectiveness of plus slides is one year from its manufacture date.

Also, less time in PFA will help. Good luck.

Calum Bonthron Did you figure out a solution? I'm having the same issue, and using what appears to be the same protocol:

Using superfrost plus gold slides, I

- freeze at -80ºC in OCT

- Section at 40um with cryostat (also using a small dab of PBS to attract them to the slide)

- Dry at RT for ~30 minutes

- freeze at -80ºC until use

- Upon use, heat on heat plate at 37ºC for 2 hours

Yet my spinal sections are still lifting off!

These are some of the traditional ways of improving section adhesion. http://www.ihcworld.com/_technical_tips/prevent_section_fall.htm

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