[Histonet] axolotl lymphatics

[Jason Palmer]

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. 

Jason 

----- 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! 

John Kiernan 
Anatomy, UWO, London, Canada 
= = = 
On 26/07/15, Jason Palmer via Histonet <histonet at lists.utsouthwestern.edu> wrote: 



Hi all, 

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? 

Thanks, 

Jason 

-- 

Jason Palmer 
Histology Laboratory Coordinator 
O'Brien Institute / St Vincent's Institute 
42 Fitzroy St, Fitzroy Victoria 3065 
Australia 
tel +61 3 9288 4045 
fax +61 3 9416 0926 
email: jpalmer at svi.edu.au 

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