[Histonet] cryojane tap transfer system

[Nicole Collette]

Hello, Richard,

I have done some sectioning of undecalcified bone with the Cryojane 
(hallelujah! it works! to paraphrase Ms. Callis) and one thing is 
that the sections won't cross-link if the tapes or the slides are 
being stored in the presence of UV light (then they are 
pre-crosslinked!), keep them dark and dry when not using, and don't 
take them into and out of the cryostat, only bring in what you need. 
I have also found that thicker sections don't crosslink as well, I've 
had trouble with sections over 6um (although I am far from am expert 
at cryosectioning). Temperature is also somewhat of a factor, I've 
found that I would section at colder temperature than I normally 
would for that tissue to get it to crosslink well- sounds 
counterintuitive, but there it is. This is purely based on my own 
trial and error, take it with a grain of salt and good luck.

Sincerely,
Nicole Collette
LLNL/UC Berkeley


At 9:01 AM -0500 11/23/09, Richard Pattison wrote:
>Hi Everybody,
>I was hoping to get some advice - I'm cryosectioning plant tissues 
>and transferring sections to slides using the Cryojane system. 
>However, i'm having problems in transferring the sections without 
>them falling apart during the tape transfer. I'm fixing my tissue 
>for 24 hours in ethanol:acetic acid (3:1), embedding in O.C.T, 
>snap-freezing and then sectioning at between 2 and 14 microns. The 
>sections seem to be ok but whenever i remove the adhesive tape from 
>the slide a large part of the tissue is removed with it. As a result 
>I lose the majority of my section. I've tried using both 1x and 1/2x 
>slides (CFSA adhesive slides from instrumedics) but neither have 
>given satisfactory results.
>Does anyone have any suggestions as to how i could reduce the loss 
>of tissue? Any advice would be much appreciated.
>Thanks
>Richard
>
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