Bone processing

From:"Marshall, Sharon,"

Dear Gayle,
I am still trying to perfect my  bone/tendon sample processing and 
cutting etc. I am getting there thanks to your help. I just have a 
few more queries. Hope it's ok. 
I want to set up a vacuum infiltration system in my lab. I suppose 
any vacuum dessicator will work?  I would probably need a glass one 
to put it in the 58 degree oven.  Is a Wheaton O ring dessicator  
special in any way? I have a glass dessicator in mind and this has a 
silicon ring.
I will follow the times you recommended for processing as the pieces 
of bone are quite large.  I am going to process as follows: 70% 
ethanol x2 11/2 hrs each, 90% ethanol x2 11/2 hrs each, 100% 
ethanol x3 11/2hrs each,  xylol x2 11/2 hrs each, histosec wax x2( at 
60degrees) 11/2 hrs each.   (all hopefully under vacuum) Does this 
sound ok.?
When you say drain and then dry the sections flat at 40C for several 
days how many days is several. ? Why does drying them flat help and 
why not at 60C ? I normally drain them and then put them on their 
sides in a rack in the 60C oven overnight. I use aptes coated slides 
as opposed to gelatin.

 I am at a lost as to which Hematoxylin to use. I usually use 
Mayer's routinely but I see you do not recommend it.  I used Mayer's 
with a celestine blue treatment prior to staining and it was fine. It 
could be darker though. Erlich's takes to long to ripen.  I probably 
will not be able to purchase Richard Allan Hx. 1 here.   I will try 
Gill II , I must find the method to make it on the internet. 

I really hope you do not mind this long discussion. 
Thanks so much for you help. 
Sharon Marshall












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