[Histonet] Problem with frozen sections for LCM

[Joel Reichensperger]

Hi everyone,

We are having an issue with ice artifact in our sections of fresh frozen 
mouse brain for use with LCM (laser capture microdissection). Currently, 
we remove the brain and immediately place it in liquid nitrogen and then 
store it at -80 until we are ready to section it. We cut 10um sections 
and once the sections are placed on the slide, they are put in a slide 
box that is kept on dry ice until all the slides are cut. At this point 
the box is placed back into the -80 until we are ready to stain them. 
Once ready to do the stain (Cresyl violet), I remove the slide and place 
it in ice cold acetone for 5 minutes to fix the sections. Should we be 
fixing the sections right after we cut them and before we put them in 
the freezer?  The outer regions of the brain are the areas that look 
terrible, while the center morphology looks nearly perfect. 
Unfortunately, we are interested in the basal forebrain. Any help would 
be appreciated.

Thanks

-- 
Joel Reichensperger
Researcher II
Southern Illinois University School of Medicine
(217)545-7292  Phone
(217)545-0145  Fax