[Histonet] freezing mouse eyes

[Danielle Crippen]

Dear all,

I have several sets of mouse eyes from perfused mice which have been in 4% PFA over the weekend.  The investigator has just informed me this morning that she needs these eyes frozen.  Can anyone with experience please elucidate what you would do in this scenario??  Should I cryoprotect?  If so, should I use a sucrose gradient: 10%, 20%, 30%??  Also, how long should each of these steps be??  At our institution, the most common freezing method is by floating the tissue in a foil boat in liquid nitrogen.  Any better suggestions??

Many many thanks to all the experts who I'm hoping will respond:-)

Cheers,

Danielle Crippen