[Histonet] RE: Re: Frozen brains on cryostat

[Thurby, Christina]

Andrea,
A couple of suggestions for you to help with sectioning the brain tissue.  If you haven't already adjusted the cryostat temperature that is a place to start.  Brains section really well at -10 to -15 C.  Make sure to let the blocks equilibrate to the temperature in the cryostat.  If you've taken the frozen blocks out of a -70 C freezer, let them sit in the cryostat for at  least 20-30 minutes to 'warm up' to the cryostat temperature.  At times, I have really good luck getting nice sections when I do the following:
1.  Shave (or face off) the block until you're ready to get a section.
2.  Using a gloved hand, place your thumb over the tissue for a few seconds.
3.  Begin sectioning - discard the first section that comes off, the second section is usually really nice (be ready to use your chilled brush) and gently pull the section toward you as you are moving the wheel (manual instrument).
4.  This method can be repeated several times - just be patient.  It does take some practice.

Finally, if you have any trouble getting the brain sections to stay adhered to the glass slides, depending on the downstream application you may want to try using the Gold Plus Slides (I think they are from Thermo Shandon or maybe Erie Scientific??)  These slides are really nice to use when working with brain tissue if you experience 'tissue lifting' during staining applications - but in our experience they can't be used for LCM work.
Good Luck!
Kristie

Christina Thurby
Bristol Myers Squibb
812-307-2093

>> On Tue, May 1, 2012 at 12:53 PM, Andrea Ferullo (non-Celgene) <
>> aferullo <@t> celgene.com> wrote:
>>
>>> Hello everyone,
>>>
>>> I recently received rat brain samples that were frozen in liquid nitrogen
>>> and embedded in OCT.  I sectioned them on the cryostat and they are coming
>>> out very wrinkled, no matter what technique I use to pick them up.  I would
>>> appreciate any tips/tricks  that anyone has to offer.  Forebrain sections
>>> are ok, but mid, hind-brain, and cerebellum are giving me a very hard time.
>>> Thanks and I look forward to your advice.
>>>
>>> Andrea


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