Re: about laser microdissection on paraffin-embeded sections
Ideally, all of your solutions used in the production of your slides
should be "RNase free". This means that you would probably want to use
millipore, DEPC-treated, autoclaved water when making solutions.
Further, and this is an unusual but desirable condition, you would
separate your regular specimens from your research (LCM) specimens.
This would optimally require a second tissue processor that is cleaned
and "sterilized", ie. RNase-free. The solutions should be as fresh as
possible and only your work load can determine how often to rotate them.
Next, all glassware and containers during the production of your of your
slides must be RNase free. Glassware, including slides, should be oven
baked and handled with gloves at all times. After mounting, always keep
the slides well protected. RNAZap
comes in handy when the containers cannot be baked. Cut and mount your
sections the day before you are going to use the LCM. The same day
would be ideal but that may be asking too much from the techs.
Typically, we mounted and stained (H&E) in the afternoon and performed
LCM the next morning.
These are some of the suggestions I can think of off the top of my head
to help you control the RNases. Not comprehensive but a beginning
Yichao WU, Ph.D candidate queried:
>Another problem I care about mostly is how would the process of making
>paraffin-embeded sections destroy RNA? I believe formalin fixation
>would protect RNA,but How about other steps? From the incubation at
60C >after sticking to slides to the deparaffinization.
"Biology in a new form..."
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