Adherence of cryosections on slides

DVR

New member
Nov 16, 2015
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#1
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

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Nov 16, 2015
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#2
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

New member
Nov 16, 2015
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#3
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.
 
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Guest
#4
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




 

sumitnbrc

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Nov 16, 2015
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#6
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