[Histonet] Undecalcified bone in MMA and antigen retrieval (Neil Hand?)

[Benjamin PERRON]

Hi all,

I'm encountering problems while trying to do antigen retrieval (AR) on undecalcified bone (rat jaw) sections embedded in methylmetracrylate (MMA). Sections fall off the slides while heated.
Section are 4µm thick, deposit on APES or gelatine coated slides, and heating is perform after total deplastification and rehydratation (I tried microwaves and warming bath in citrate buffer pH=6.0).

Could some around here share theyre experience with AR on undecalcified bone sections and in particular tell me :

-does section thickness as an impact on sections detachment while heating (I know some are performing thinner section, but 3-4µm is the thinner we are (in routine) able to do with rat jaw)
-is theyre a particular coating for slides that would better fit that what we are using (APES or gelatine). Someone suggest me Poly-l-lysine.

All tips and advices from experimented in this field would be very helpfull.

Benjamin Perron
Student (Master in Cell Biology)
EA2496
Montrouge
France