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

[Jack Ratliff]

Benjamin,
 
How about trying Haupt's coated slides. I am not sure if these slides are now commercially available, but you can coat them yourself. In the past I have made the solution by scratch and also used the commercially prepared solution from Fisher. When you go to the Fisher Scientific website, the product will be listed under Harleco Gelatin Fixative #785-71 @ 500 mL.
 
This is a concentrate solution, so it will be best to aliquot a small amount into a beaker, gently warm in a lab use only microwave for 10-20 seconds or a hot plate until the solution is in a complete liquid state, freely flowing. Then cut it 1:1 with 50% EtOH to prepare the working solution. Place 1-2 smal drops on a slide and use another slide of the same width to gently move the solution across the specimen mounting area. Finally, let it dry flat for 5-10 minutes and then you are ready to use.
 
Let me know if you need anymore help!
 
Jack Ratliff
 
> From: perron.b <@t> wanadoo.fr> To: histonet <@t> lists.utsouthwestern.edu> Date: Mon, 12 Jan 2009 19:02:05 +0100> Subject: [Histonet] Undecalcified bone in MMA and antigen retrieval (Neil Hand?)> > 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> _______________________________________________> Histonet mailing list> Histonet <@t> lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet