[Histonet] sucrose sectioning

[RCHIOVETTI <@t> aol.com]

In a message dated 2/20/2006 1:26:21 PM US Mountain Standard Time, 
sjchtascp <@t> yahoo.com writes:

> I work with a scientist that never seems pleased with my cryosectioning of 
> fixed sucrose infiltrated liver. 

Hi Steve,

I'm most familiar with using sucrose as a cryoprotectant / infiltration 
medium when doing cryoultramicrotomy and immunolabeling at the EM level.  In those 
cases, sucrose is normally used at around 2.3M concentration after light 
aldehyde fixation, and it usually cuts well at around -90 C (the cryosectioning 
kits for ultramicrotomes use liquid nitrogen, and they allow you to cut at very 
low temps).

I never could get sucrose to cut well above about -60 to -65 C.  It just 
turned to mush and became a sticky, stringy mess.

A normal cryostat w/ independent specimen cooling should get down to about 
-50 C, but I'd think you'd be right on the edge of being able to use sucrose.

Unless there's some reason not to use the stuff, maybe you could try O.C.T. 
or a similar cryo medium designed to be firm at higher temps (say round -20 to 
-30 C).

Good luck!

Cheers,

Bob



Robert (Bob) Chiovetti, Ph.D.
The Microscope Works
Arizona's Microscopy Resource
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