[Histonet] Sialidase digestion [long]

[Smith, Allen]

  Many years ago, I used sialidase and a control, followed by colloidal
iron, to study primate salivary glands (A.A. Smith: Major salivary glands of
the Philippine tarsier.  Folia Primatologica 10: 113-130, 1969). 
  I added 2 ml of reconstituted cholera filtrate (4000 microgram units of
sialidase) to 41 ml of 0.05 M pH 5.5 acetate buffer (36 ml 0.05 ml sodium
acetate adjusted to pH 5.5 with 0.1 M acetic acid plus 324 mg sodium
chloride and 48 mg calcium chloride dehydrate).  The control was the buffer
without the cholera filtrate.  I incubated for 24 hours at 37 degrees C.
  Audrea Vaughan (former graduate student, now teaching at Miami-Dade
College) used sialidase and colloidal iron to study rhesus monkey salivary
glands.  She used 1.0 micromolar units of Clostridium perfringens sialidase
(Sigma type V) in 40 ml of 0.1 M pH 5.5 acetate buffer.  She incubated 24
hours at 37 C.
  Both of us used Mowry's colloidal iron ( R.W. Mowry: J. Clin. Invest. 7:
566-576, 1958).  The procedure is given in Humason's ANIMAL TISSUE METHODS
(pp. 301-303 in my copy of the 4th edition).  I have also attached it here. 
Colloidal iron is a more sensitive detector of sialomucins than alcian blue
or zirconyl hematoxylin.
  Audrea Vaughan found that sialidase worked well on tissues fixed in
formalin, alcohol-formalin, or Carnoy's.  It did not work on tissue fixed in
Zenker's or Helly's fluid.  Even the buffer removed the sialomucins from
tissues fixed in Bouin's fluid.

  Allen A. Smith, Ph.D.
Professor of Anatomy
Barry University School of Graduate Medical Sciences
    Podiatric Medicine and Surgery
Miami Shores, Florida  33161


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Laurie
Colbert
Sent: Wednesday, June 08, 2005 4:23 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Sialidase digestion

Has anyone ever done (or heard of) a colloidal iron stain with sialidase
digestion?  Our pathologists have a mucinous skin tumor and are trying to
differentiate between a primary breast CA and a primary eyelid tumor.  It
may be run in conjunction with hyaluronidase digestion.  Any references or
procedures??

Laurie Colbert
Huntington Hospital
(626) 397-8620  

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