[Histonet] Bone processing
gcallis <@t> montana.edu
Thu Jul 28 16:14:36 CDT 2005
Those protocols are from people who did their animal tissues in a certain
way, and not everyone does it just exactly like the protocol submitted for
the Animal Processing Manual - the protocols are not written in stone, but
are good guidelines for successful animal tissue work.
As for our mouse bone work, we use a Sakura Finetek VIP, and extend
processing for mouse joints (denser due to the collagen matrix of
decalcified bone). We do ambient i.e RT temperatures for dehydration and
clearing and use vacuum and pressure throughout. Adding temperatures to
alcohols, clearing agents sometimes adds to hardness of tissue at
sectioning - animal tissues, in particular the tiny mouse soft tissues. In
general, animal tissues are much leaner, less fatty than human tissue and
overdehydration tends to dry out rodent tissue more. This species is not
the only one that suffers from lean i.e birds, reptiles, rabbit, and many
others. We tend to do custom processing for rodent work, and have
special schedules for mouse brain versus hamster brain versus rat brain
versus lung and other soft tissues. Larger animals processing can be more
like human tissue processing i.e. cow, horse, dog, cat, but sometimes these
can be dry also.
What you want to do is remove the free water from the tissue spaces and not
the bound water found on the proteins (that is what makes mouse spleen hard
little nuggets sometimes referred to as overprocessing). If your tissue
seem dry at sectioning - you can soak a faced block on ice with water on
top and reevaluate if you need to cut down on total time of processing, in
small increments of time however.
At 02:19 PM 7/28/2005, you wrote:
>New to research, just received the "Animal Processing Manual". I'm about
>to process mouse joints and I noticed a much longer time especially in the
>Along these lines I also noticed processing for mouse/rat tissue calls for
>only RT processing and no vacuum/pressure..
>I have a clinical background so some or allot of this is a "tad"
>new. Anyone up to explaining the big picture difference between human
>tissue verses animal tissue processing.
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