Thank John very much. I will check out your references. Hope we are not only be able to follow the protocol, but fully understand it.
Dorothy Sent from my iPad > On Jan 16, 2014, at 1:37 PM, John Kiernan <jkier...@uwo.ca> wrote: > > Methacrylate monomers are sold with added hydroquinone to inhibit spontaneous > polymerization. The NaOH treatment to remove hydroquinone dates from the > earliest years of plastic embedding (Newman et al. 1949 Science 110: 66-68). > The NaOH treatment is not necessary, however, because the stabilizing action > of hydroquinone can be overcome by simply using a larger amount of the > polymerization catalyst. About 2% of either benzoyl peroxide or > 1,2-dichlorobenzoyl peroxide will catalyze the polymerization of stabilized > methacrylate monomer. See Hayat, MA 1981 Principles and Techniques of > Electron Microscopy Vol 1. Baltimore: University Park Press, pp. 167-168. > > See Baker, JR 1960 Cytological Technique, 4th ed. London: Methuen, pp.77-84 > for a thorough but simple explanation of the chemistry of methacrylate > embedding. The whole book (which is a classic) is available, free, at > http://archive.org. > > John Kiernan > Anatomy & Cell Biology > University of Western Ontario > London, Canada > = = = >> On 16/01/14, Dorothy Hu <abt...@gmail.com> wrote: >> Thanks Jack's answer. I though I shouldn't to destabilization step as well. >> But my boss wants to do it. Is there a chemical theory behind this? Why >> previous protocol has this step (it must be reason)? And why the step is >> skipped now? I wish I know before approaching my boss. >> >> Additionally, there is a problem always bothering me in plastic sectioning. >> I often get cortical bone shattering ( crumbly) in the midshaft of long >> bone. Cortical bone and marrow are operate in the midshaft. I don't know >> why is that. It's not happen in the two ends of tibia and femur bone. And >> doesn't happen on ankle bones. Is that because the bone was stored in 70% >> EtOH too long (~one year)? I think infiltration is good since I did three >> times, 2 days each infiltration under vacuum condition in 4oC. If I use 40% >> EtOH to fix and store the bones will help? >> >> One more question. What is advantage if I use perkadox 16 to replace BP as >> catalyst? Do I have to dry MMA//DBP/perkadox 16 mixture through CaCl2? >> >> Thanks in advance for your help. >> >> Dorothy >> MGH endocrine histocore >> >> >> On Wed, Jan 15, 2014 at 11:02 PM, <abt...@gmail.com> wrote: >> >> > I am new to MMA plastic bone technique. Some one gave me his protocol, in >> > which has NaOH and d-water to wash MMA mixture before drying it in CaCl2. >> > But others told me I don't need to do the destabilization step. Could any >> > expert in this area to tell me if this step is necessary? And why have to >> > do? >> > >> > Sent from my iPad >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
