[Histonet] Sections falling off

[Gerard Spoelstra]

Hi Pablo

I have found with bone sections, which I pick up with silanised slided, that if I apply gelatin to the slides just  prior to picking up the sections, the sections don't lift. The slides are dipped in the gelatin solution, then the sections are picked up off the surface of the water.
 I use roughly 0.02 of a gram of gelatin/ 100 ml water dissolved with microwave.  

The gelatin seems to alter the surface tension of the water as there is no pooling around the edges of the paraffin. The sections are then dried for a hour in a 60 degree oven, before staining.

Gerard Spoelstra
Veterinary Histology
Murdoch University
Western Australia

________________________________________
From: SANCHEZ QUINTEIRO PABLO via Histonet [histonet at lists.utsouthwestern.edu]
Sent: Thursday, March 23, 2017 8:06 AM
To: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Sections falling off

Lots of thanks Ruth! What you tell us seems very sensible.

But I am unsure about your protocol. After 24 h Bouin we transfer the brain to 70 alcohol and start embedding several days later.

Must we avoid this storage in 70 ethanol? Instead must we wash the samples from Bouin after 24 h in water and start embedding from that point?

Thanks for your input.

Pablo
________________________________
De: Yaskovich, Ruth A (NIH/NIDCR) [E] <ryaskovich at dir.nidcr.nih.gov>
Enviado: miércoles, 22 de marzo de 2017 20:58:35
Para: SANCHEZ QUINTEIRO PABLO
Asunto: RE: [Histonet] Sections falling off

Pablo,
 Don't leave them in 70 rinse the bouin's out you can rinse cassettes in running water and then put on processor. Alcohol really dries out the CNS tissues and can cause that falling off the slide. I have been there.
Good Luck
Ruth
N.I.H.

-----Original Message-----
From: Pablo Sánchez Quinteiro via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Wednesday, March 22, 2017 2:22 PM
To: histonet at lists.utsouthwestern.edu
Subject: Re: [Histonet] Sections falling off

Thanks for your reply Shirley.

After 24 h fixation in Bouin samples are passed into 70º ethanol.

I have tried drying the sections at RT for short or long period (30´) and then 37º oven overnight.

The tissue is mouse brain, sections thickness 8 microns.

Pablo



El 22/03/2017 a las 18:45, Shirley A. Powell escribió:
> Are you washing the bouins out of the tissue well before processing, that may be one source.  Also how long are you drying your sections before staining? What type of tissue are you processing?
>
> -----Original Message-----
> From: Pablo Sánchez Quinteiro via Histonet
> [mailto:histonet at lists.utsouthwestern.edu]
> Sent: Wednesday, March 22, 2017 1:39 PM
> To: histonet at lists.utsouthwestern.edu
> Subject: [Histonet] Sections falling off
>
> Dear Histonetters,
>
> I am in trouble with my paraffin sections -samples fixed in Bouin's liquid.
>
> Just after the first steps: Two xylol baths and 100% ethanol most of them are falling off. Slides are silanised.
>
> I do not think it was an adhesion problem. What do you think about?
> Embedding, cuttibg?
>
> Thanks a lot in advance
>
> Pablo
>
>
> ---
> El software de antivirus Avast ha analizado este correo electrónico en busca de virus.
> https://www.avast.com/antivirus
>
>
> _______________________________________________
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet


---
El software de antivirus Avast ha analizado este correo electrónico en busca de virus.
https://www.avast.com/antivirus


_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
Histonet mailing list
Histonet at lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet