[Histonet] Araldite
Coughlin, Beth
coughlin <@t> musc.edu
Thu Aug 18 10:10:11 CDT 2011
Anyone have advice for how to get sections from epon blocks that are very soft??
I used a rapid embedding protocol for making epon blocks that I found online.
Unfortunately, I misunderstood the instruction that I was to use araldite alone for the embedding media.
I used the kit that came with the araldite (ddsa and dmp 30).
I didn't actually use the steps for polymerizing recommended below. I just used the early steps for speeding up the processing. I just put my samples in at 60 degrees overnight, as usual.
WASH...............PBS..................2X5 MIN.
1%OsO4 in PBS...........................20 MIN RTEMP.
WASH...............PBS..................2X5 MIN
*70% ETOH...............................3 MIN
95% ETOH................................3 MIN
100% ETOH...............................2X5 MIN
PROPYLENE OXIDE.........................2X5 MIN
1:1 P.O.:ARALDITE 502...................30 MIN (REMOVE Caps)
1:3 " " ...................20 MIN
PURE ARALDITE 502.......................10 MIN
EMBED
CURE IN 75C OVEN FOR 50 MIN. PLACE IN 95C OVEN FOR 50 MIN.
PLACE IN FREEZER AND LET COOL 10 MIN.
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