Fwd: Re: [Histonet] cryostat mounting

[Rene J Buesa]

[As shared response]
Rene J Buesa <rjbuesa <@t> yahoo.com> wrote:  Date: Fri, 10 Mar 2006 05:47:20 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] cryostat mounting
To: 0104684s <Joseph.Garvin <@t> NUIGALWAY.IE>

  Hi Joseph:
  Distilled water is what I always used when doing direct immunofluorescence (DIF) in skin biopsies, either punch or shave.
  I did it in the following way:
  1- you set the holder in the thermal absorption bar in the side of the cryostat
  2- you should have at least 1 pair of fine tipped forceps also inside the cryostat so they are cold enough
  3- you hold a dropper with distilled water with your left hand and with your right hand you hold the skin biopsy with the fine tipped forceps
  4- you add a small amount of water (about 0.5 mL) on the chuck and as the water begins to solidify you set the skin sample over that drop of water that is starting to become ice
  5- once the skin is in place on the water/ice interface, you keep adding water and building a small and round ice block over it
  6- the ones I enjoyed most doing were those around the skin shave biopsies.It was like building a cocoon around them!
  7- once the skin biopsy is totally covered with the ice, you take it in the chuck to the cryostat and start sectioning.
  The sections (over a very cold permanent or steel blade) slide beautifully and when picked up with the slide (always much wamer) immediately melts out leaving the skin section on it.
  I personally prefer cutting skin biopsis for DIF when they are embedded in water, rather than in OCT ("too messy" for my preference).
  Hope this description will help you!
  René J.

0104684s <Joseph.Garvin <@t> NUIGALWAY.IE> wrote:
  Has anyone any experience of mounting frozen tissue sections onto the chuck in 
a cryostat using water as opposed to OCT

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