Fwd: Re: [Histonet] Bone tissue

[John Kiernan]

 Just for the record, Mollifex isn't alkaline. It is probably either Baker's 1941 mixture (see below) or the following (parts by volume):
    95% ethanol 952
    Glycerol 32
    Acetone 80 
    "Liquid phenol" 8
 - - -
"Liquid phenol" is probably liquefied phenol, USP, which contains about 10% water and at least 89% phenol (w/w), or something similar.
 For more information and discussion, see Maria Wynnchuk (1992) Minimizing artifacts in tissue processing: Part 1. Importance of softening agents. J. Histotechnol. 15(4): 321-323. This paper also discusses effects of softening faced blocks on H&E staining.
  
John R Baker (1941) used 9 parts 60% ethanol and 1 part glycerol (J. Roy. Microsc. Soc. 61: 75), whereas R. C. Pearlman and Buell C. Cole (1951) sang the praises of 1% dishwashing detergent and similar solutions (Stain Technol. 26(2): 115-118). Manfred Gabe (Histological Techniques, Paris, 1976, a great book) favoured soaking faced paraffin blocks in cold water. For what it's worth, I have found water helpful when sectioning decalcified rats' heads. With all these well documented and inexpensive softening liquids it should never be necessary to resort to a proprietary product whose composition is kept secret from the buyer and user.
  
 John Kiernan
 Anatomy, UWO
 London, Canada
  = = =
 ----- Original Message -----
> From: Rene J Buesa <rjbuesa <@t> yahoo.com>
> Date: Monday, February 22, 2010 16:43
> Subject: Re: [Histonet] Bone tissue
> To: histonet <@t> lists.utsouthwestern.edu, Reuel Cornelia <Reuel.Cornelia <@t> tsrh.org>
> 
> > First of all, Mollifex or any other alkaline substance will do 
> > nothing "useful" to the bone.
> > I tend to think that you processed the bone before it was 
> > completely decalcified and that is the cause for an incomplete 
> > infiltration and a subsequent difficult sectioning.
> > René J.
> > 
> > --- On Mon, 2/22/10, Reuel Cornelia <Reuel.Cornelia <@t> tsrh.org> 
> > wrote:
> > 
> > From: Reuel Cornelia <Reuel.Cornelia <@t> tsrh.org>
> > Subject: [Histonet] Bone tissue
> > To: histonet <@t> lists.utsouthwestern.edu
> > Date: Monday, February 22, 2010, 4:36 PM
> > 
> > 
> > We have a difficulty cutting metatarsal bone . It seems that our 
> > sections are so dried up. I was thinking that our dehydration 
> > have something to do with this which we have placed it in a 
> > wrong processing procedure for our large bone. The tissue is 4 
> > mm thick and 1-2 cm in length and width and  was dehydrated in 
> > 70% - 4 hrs, 80%-4 hrs,95% -4 hrs and 2 changes of 100% 3 hrs 
> > each, paraffin is 4 hrs each 2 changes. The tissue was 
> > decalcified in 14% EDTA. When we start cutting them it is so 
> > brittle and we could not even create a section. I have surfaced 
> > decal it and also place in a softener Mollifex some of it work 
> > but some does not work. Please help us remedy this tissue. Thank you.
> > 
> > 
> > 
> > 
> > Reuel Cornelia, BS MT, AMT
> > Cellular Pathology
> > Texas Scottish Rite Hospital for Children
> > 2222 Welborn Street
> > Dallas, TX 75219
> > Tel: 214-559-7766
> > fax: 214-559-7768