Grantia slides

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Kambio
Posts: 17
Joined: Mon Jul 25, 2016 7:22 am

Grantia slides

#1 Post by Kambio » Mon Jul 25, 2016 12:35 pm

Hi there.
I have had some problems putting Grantia t.s. on slides and keeping them there wholly intact. I have stained them with chlorazol black but find that they are so fragile in xylol that the movement of the DPX over the specimen when placing the c/s over the top causes some of the specimen to 'float' away. Any tips out there??

Peter
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Joined: Sun Oct 12, 2014 5:34 pm

Re: Grantia slides

#2 Post by Peter » Mon Jul 25, 2016 7:17 pm

Hi Kambio,
If you give us some more information on your microtechnique we may be more able to help, (eg how are you flattening your sections, what are you using to adhere them to the slide).
Peter.

Kambio
Posts: 17
Joined: Mon Jul 25, 2016 7:22 am

Re: Grantia slides

#3 Post by Kambio » Mon Jul 25, 2016 9:34 pm

Hi Peter.
I use a microtone to cut the sections at about 10um and adhere the cut section to the slide with a albumin type product. Then they are allowed to 'dry' a RT for a few days before placing them through a series of solutions.....etc

Peter
Posts: 226
Joined: Sun Oct 12, 2014 5:34 pm

Re: Grantia slides

#4 Post by Peter » Tue Jul 26, 2016 9:10 pm

Hi Kambio,
I have never sectioned these specimens, however; I am assuming you are cutting paraffin sections and flattening them by warming them while floating them on water on a slide. If this is so, I would drain the water from the slide, place it on a solid bench, cover with a piece of saturated filter paper (I use Whatmans No. 50) and roll over the slide with a rubber roller, peel off the paper, dry the slide, and process with your usual reagents.
Are you using any reagents which may soften/dissolve albumin?
Hope this helps.
Peter.

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mrsonchus
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Re: Grantia slides

#5 Post by mrsonchus » Tue Jul 26, 2016 11:46 pm

Kambio wrote:Hi Peter.
I use a microtone to cut the sections at about 10um and adhere the cut section to the slide with a albumin type product. Then they are allowed to 'dry' a RT for a few days before placing them through a series of solutions.....etc
Hi Kambio, can you be specific please, are you sectioning them within (i.e. infiltrated with and embedded in) wax blocks and if so are you using a rotary microtome?
Your problem is one that I have much experience of in my earlier days learning my technique but have now completely eradicated, but I need to know exactly how you are sectioning, placing onto slides and drying your sections to help old chap.

The more details the better. :)

p.s. including timing wherever water is involved.
John B

Kambio
Posts: 17
Joined: Mon Jul 25, 2016 7:22 am

Re: Grantia slides

#6 Post by Kambio » Thu Jul 28, 2016 7:00 am

Hi John B.
Thank you for your input.

I use a rotary microtone to cut the wax embedded specimen at about 10um. sections. Then place a cut section onto a slide that has a thin smear of an albumin product and a 3% formalin solution to 'maneouver' the section in place. I chose chlorazol black in 70% alcohol to stain the dried sections as this didn't involve as many 'steps' as the H & E staining method. After a few days of the sections 'drying onto the slide I put the rack of sections into the usual de-waxing and hydrating solutions until they were in 70% alcohol then I stained them in the chlorazol black. The next steps involved dehydrating to Xylol ready to mount in DPX. This methodolgy is a basic process that I have used numerous times for other specimens but these Grantia sections were trickier than I expected.

Kambio
Posts: 17
Joined: Mon Jul 25, 2016 7:22 am

Re: Grantia slides

#7 Post by Kambio » Thu Jul 28, 2016 7:02 am

Hi Peter.

Thank you for your reply. You have given me some techniques which I will incorporate into my methodology for the next batch of grantia, I will let you know how they go.

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