[Histonet] lymphatics

[anh2006 <@t> med.cornell.edu]

I read Dr. Kiernan's post with much interest. Has anyone on Histonet had
direct experience with these protocols? I would love to get some first
hand experience feedback before I try it out.

Thanks,
Andrea


> Dear Hal Hawkins,
>
> The endothelium of lymphatic vessels can be stained by
> enzyme activity histochemistry. The enzyme is a
> 5-nucleotidase (belongs to the phosphatase family).
> Ordinarily this type of method is carried out with frozen
> sections of tissue that has been minimally fixed in
> formaldehyde or glutaraldehyde, or with unfixed cryostat
> sections that have been briefly fixed in cold acetone. The
> enzyme in lymphatics may be fairly robust because its
> activity can survive embedding in glycol methacrylate.
>
> The references below may help. If you choose one of these
> methods it will be important to consult the original
> publication because there's more than one sort of
> 5-nucleotidase. Each ref is followed by brief notes that I
> took when reading the papers. [An R in the acquisition
> number means I've got a Reprint of the whole paper; an A
> usually means I have a copy of the paper's Abstract. No
> letter with the acquisition number usually means I've seen
> the whole paper and took notes in the library.]
>
> 8194R. Kato,S; Yasunaga,A; Uchida,U (1991):
> Enzyme-histochemical method for identification of lymphatic
> capillaries. Lymphology 24, 125-129.
>   Glycol methacrylate-embedded sections stained for
> 5'-nucleotidase & alkaline phosphatase. 5-N-ase in lymph
> capills; AlkP-ase in blood capills.
>
> 9239R. Nishida,S; Ohkuma,M (1993): Enzyme-histochemical
> staining of dermal lymphatic capillaries by guanylate
> cyclase. Lymphology 26, 195-199.
>   Enzyme histochemical staining distinguishes lymph from
> blood capillaries. Says 5-nucleotidase & adenylate cyclase
> do so too. (Used unfixed cryostat sections.)
>
> 9665R. Okada,E (1994): An improved enzyme-histochemical
> method for identification of lymphatic capillaries on
> paraffin sections. Lymphology 27, Suppl, 732-735.
>   Staining of blood & lymph capillaries. Enzyme
> histochemistry for 5-nucleotidase in presence of levamisole
> for lymph capills; alkaline phosphatase for blood capills.
>
> 11114A. Miura,M; Kato,S; von Ludinghausen,M (1998):
> Lymphatic drainage of the cerebrospinal fluid from monkey
> spinal meninges with special reference to the distribution
> of the epidural lymphatics. Arch. Histol. Cytol. 61(3, Aug),
> 277-286.
>    5'-nucleotidase staining for lymphatics; alk phosphatase
> for blood capillaries (Kato et al '91,'93). Also traced
> carbon particles from cisterna magna. Lymphatics and carbon
> found on surfaces of cervical & thoracic (most at brachial
> plexus levels) roots; not lumbosacral. Epidural lymphatics
> most developed on dorsal surface of lower cervical dura.
>
> I hope this helps.
> --
> -------------------------------
> John A. Kiernan
> Department of Anatomy and Cell Biology
> The University of Western Ontario
> London,   Canada   N6A 5C1
>    kiernan[AT]uwo.ca
>    http://publish.uwo.ca/~jkiernan/
>    http://instruct.uwo.ca/anatomy/530/index.htm