[Histonet] axolotl lymphatics
jpalmer at svi.edu.au
Fri Jul 31 00:07:36 CDT 2015
Thankyou for your reply John. There is no-one whose suggestions I value more on Histonet, and your book (H and H methods) is also my go-to source for all things staining-related.
I didn't mention in my post that the material I am working with is paraffin-embedded, so will not be any good for enzyme demonstration. If I can manage to get some tissue for freezing at some point then I will consider your suggestions. From memory, I tried something similar 20-odd years back on frozens, before the advent of podoplanin, LYVE-1 etc. immunostaining for lymphatics on FFPE, but without much success. Might be worth a re-visit.
----- Original Message -----
From: "John Kiernan" <jkiernan at uwo.ca>
To: "Jason Palmer" <jpalmer at svi.edu.au>, histonet at lists.utsouthwestern.edu
Sent: Wednesday, 29 July, 2015 2:06:29 AM
Subject: Re: [Histonet] axolotl lymphatics
Instead of an antibody, you might consider enzyme activity histochemistry, which is much less expensive.
Demonstration of 5-nucleotidase activity in the presence of levamisole detects lymphatic endothelium. Sections can also be stained for alkaline phosphatase activity in the endothelium of blood vessels. Here are a few references.
Kato, S., Yasunaga, A. and Uchida, U. (1991). Enzyme-histochemical method for identification of lymphatic capillaries. Lymphology 24 :125-129.
Ohkuma, M. (1994). Simultaneous double staining for the blood and lymphatic capillary. Lymphology 27 , Suppl:106-107.
Okada, E. (1994). An improved enzyme-histochemical method for identification of lymphatic capillaries on paraffin sections. Lymphology 27 , Suppl:732-735.
Ji, R.C. and Kato, S. (2003). Lymphatic network and lymphangiogenesis in the gastric wall. Journal of Histochemistry and Cytochemistry 51 :331-338.
Needless to say, none of these relate to amphibian tissues!
Anatomy, UWO, London, Canada
= = =
On 26/07/15, Jason Palmer via Histonet <histonet at lists.utsouthwestern.edu> wrote:
I need to find an antibody that will label lymphatic endothelial cells in the axolotl. Does anybody have any experience or ideas? I have tried a couple of our anti-mouse and anti-human Abs for podoplanin and LYVE-1 but no cross-reactivity so far. I have no experience with staining of non-mammalian tissues - maybe an anti-frog Ab would cross react? Does anyone have experience with other amphibians?
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