Re: [Histonet] Problem with frozen sections for LCM

From:"John A. Kiernan"

The cause of your trouble is putting the specimen
into liquid nitrogen, which forms an insulating
blanket of nitrogen gas around the immersed
object. Try putting your hand in the liquid N2,
and see how long it takes to freeze it solid. :)

The archives at contain many
Histonet hints about how to freeze tissue. Yours
is a question that is frequently asked. 
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
Joel Reichensperger wrote:
> 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
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