Adherence of cryosections on slides

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DVR
DVR's picture
Adherence of cryosections on slides

Hi,
I am working with intestinal tissue from rat. When I try to stain the tissue sections after taking them on to the slides. They come off from the slides. I have used ultrafrost, superfrost and silanated slides. But none of them could hol the tissue until I finish the staining process. Are there any other kinds of coated slides or any methods to hold the tissues on the slides.
I will be very thankful for your help.

Venkat

R Bishop
R Bishop's picture
Venkat,

Venkat,

I had similar problem with liver cryosections.  Turned out I was not treating the slides properly during the staining procedure.  

Check here for the protocol that got me passed the problem, which was gentle fix just before staining.

Bishop

DVR
DVR's picture
Thank you very much Bishop!

Thank you very much Bishop!
I have tried the paraformaldehyde fixation but it doesn't help me. I think I can try with acetone. I have been working on this for the past one month and its very frustrating when the sections don't stay on the slides.

Guest (not verified)
Guest's picture
 Hi I work with the brain

 Hi I work with the brain that has a high fat content that also tend to fall off.  I haven't found any commercial  slides that work.
This is my solution:
1) take ordinary uncoated slides. put them in slide racks, use gloves to stop grease getting on the slides.
2) dip them in 1% gelatin (heat the gelatin in distilled water, use the microwave, less messy)
3) dry or put in incubator for 6 hours. Store room temp until used.

4) Place section on the slide.
5) Dip the section in fixative (4% para) for 30 sec.  This fixes the section to the gelatin and hardens the gelatin. The 30 sec fixation has not decreased antigenicity for the AB I use.
Good luck
OCT

R Bishop
R Bishop's picture
Nice.  Thanks OCT!  I am

Nice.  Thanks OCT!  I am definitely gonna try that gelatin/paraformaldehyde trick next time.

Bishop

sumitnbrc
sumitnbrc's picture
You can  coat slides with  3

You can  coat slides with  3-AMINOPROPYLTRIETHOXYSILANE
Preparation Instructions

3-aminopropyltriethoxysilane (APES) is soluble in
toluene  and acetone.

APES is soluble in ethanol at 100 mg/mL, soluble in
chloroform and soluble in acetone (see below).
Stability in solvents with hydroxyl groups (alcohols,
water) is very poor, and only limited in acetone.

Storage/Stability

If stored as recommended this product will have a shelflife
of two years.

Procedure:

Slides should be "soaked" in methanol or acetone for
about two minutes, then air-dried. This assures that the
slides are free of any trace of oil or water.
1. Mix 10 mL of the silane with 500 mL acetone. (This
makes a 2% solution, which is stable for 8 hours,
after which the color will change.)
2. Dip clean dry slides in silane solution for 2 minutes.
3. Wash the slides in 2 changes of distilled water.
4. Dry completely in oven and cool. Autoclaving, if
    desired, will not harm the coating.
5. Store treated slides in boxes at room temperature

www.sigmaaldrich.com/etc/medialib/docs/Sigma-Aldrich/Product_Information_Sheet/a3648pis.Par.0001.File.tmp/a3648pis.pdf

ncwildb88
ncwildb88's picture
Dear OCT,

Dear OCT,

would the protocol you described work with acetone instead of 4% para?