[Histonet] Sections falling off

[Pablo Sánchez Quinteiro]

Thanks a lot Shirley,

How long you wash in water the tissue before embedding? Where does the 
tissue come from? 70 alcohol?

Pablo



El 22/03/2017 a las 20:06, Shirley A. Powell escribió:
> I cut autopsy material for the medical examiners here and they are usually very bloody and thick sections.  I face my blocks, soak them in an ice slush for 30 min to an hour or on a moist towel in the freezer overnight.  I section them at 5 microns, a little thinner than yours.  I use a product called Sta-On from Leica, it was a Surgipath product but Leica bought them out.  I use a 10 ml to 990 ml water in my water bath.  Charged slides or coated ones do not hold the bloody tissues on as well, so I use plain slides.  After sectioning I stand them up for all the water to drain from between the section and the slide.  Next I place on a hot plate/slide warmer at a temp just above the melting point of my paraffin to help the section relax and smooth out the micro folds that may be in the section. This takes s just a few minutes to melt the paraffin from around and in the section.  I then place in the slide staining rack and place in an hot air slide dryer for at least an hour, but for me, I leave them overnight at 60 degrees.  The next morning I stain my sections.
>
> Brain is a tissue that does not adhere as well as others to the slide so when staining be very gentle, agitate them in each solution but I would not agitate vigorously.
>
> The reason I asked about washing the bouin's out, is that it could change the charge on your slides and affect the stain.  I was always taught to wash the tissue before beginning the dehydration in the processing so that residual picric acid would not affect my staining as well.
>
> Hope this makes sense and helps.
>
> Shirley
>
> -----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
>>
>>
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