[Histonet] Sucrose infiltration and Bone question...

[Jamie E Erickson]

Hi All,
             I have two question about sucrose infiltration....First,  I 
have infiltrated many mouse brain with sucrose 30% in the past and it has 
always been at 4degrees C (dc) and I'm asking myself now  why? My mouse 
brain are fixed in paraformaldehyde or formaldehyde for 24 hours prior so 
why sucrose @ 4dc? Would it not infiltrate better (faster)  at room temp? 
I could not find out why 4dc...

My real problem is that  I am tiring to get good mouse tibia/femur  frozen 
sections and I am using D-profile blade and a tape transfer method. It 
worked great on my rats sections but not mouse so I think the sucrose is 
not getting in to the bone.   I can't use the sinking tissue as an 
indicator because the bone sinks right away.

Should I go longer in sucrose (maybe 48 Hrs)  but at 4dc or room temp  ?

Would adding 2 % DSMO to sucrose facilitate penetration..??

Any suggestions would be great..

Thanks have a great day..

Jamie
_______________________________
Jamie Erickson
Sr. Research Associate II M.S. HTL (ASCP)
Department: DSMP
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
508-688-3134
FAX: 508-793-4895
e-mail: jamie.erickson <@t> abbott.com