Re: [Histonet] HELP, Plastic sectioning

From:Jorge Tornero

Hi,

I think the first thing you have to change is your knife. As far as I
know, it must be a tungste-carbide knife (or glass).

I use a leica rm-2145 microtome whit a D-profile knife. I cut with an
angle of about 6 degrees.

But also I have suffered from shattering and I was annoyed because we
even use a leica EM-TP tissue proccessor to do the embedding. We use a
very long protocol for dehydration and embedding (24 EtOH 70, 24 h
EtOH 96, 24 h EtOH 100, 48 h 1:1 and 60 hours of pure activated resin)
and it worked for everybody except us. Why? One day I realized that we
put the pure resin and the 1:1 solution at the begginning of the
proccess (when we started the EM-TP) and oviously the resin was
damaged (it turns with a brownish color) when its turn in the
processor began. So now, we put the resin into the processor hours
before its use (because it is friday and we dont work on weekend and
the dwarf inside the processor works for us and changes the liquids)
And now everything is fine. Check if there is a failure in your
protocol or equipment and remember to store any resin or its solutions
in the fridge at 4%. Our problem was caused because the EM-TP only
refrigerates the vial that is being used in that moment. So we trust
that we had 4 ºC in our processor but it only was in the vial that was
running.

Saludos,

Jorge Tornero
Instituto Español de Oceanografia
Cádiz - SPAIN

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