Ian,

When I did marine fish in the past, I used 0.2 micron filtered 
seawater for my buffer when making the Karnovsky's. This was for SEM, 
but the cilia of the lateral line sensory end organs looked fine, as 
did the surfaces of the sensory cells.
But! Let the water warm up to 25 deg C or so -- cold seawater will 
precipitate the formalin and probably also the glutaraldehyde.

Phil

Adrian,
	Measure Sorensen's standard buffer with an osmometer, 
although physiological it does have a high osmolarity, that's why I 
thought it might work. Yes, I've started trawling through the 
journals etc. looking for the "standard" sea water substitute. As for 
the Clyde, well, if the cold and rain don't get me the pollution 
certainly would.
Ian.

Date: Mon, 03 Dec 2001 19:27:20 -0400
From: nina leek 
X-Mailer: Mozilla 4.04C-MOENE (Macintosh; U; PPC)
To: "Dr. Ian Montgomery" ,
    histonet@pathology.swmed.edu
Subject: Re: Fixation buffer.

Ian:
Sorensen buffers are physiological strength aren't they, and so 
somewhat hypotonic to sea-water?  There are a number of recipes for 
artificial sea waters in older biochemistry books, and I would have 
thought any of them would work as a base for making up the fixative. 
I may be able to dig some out if you need me to.  In this weather, it 
would be more comfortable than going down to the Clyde to dip a 
bucket in.
Good luck,

Adrian Leek.


Dr. Ian Montgomery wrote:

  	Preparing fish larvae (sea) for EM, any suggestions for a 
suitable buffering system given that the levels of salt in sea fish 
are  ~3 times that of mammals. Do I simply make up the fixatives 
using sea water? Sorenson buffered fixatives have a high osmolarity 
that might be another buffer. Time, unfortunately, is short so I 
can't run a series of trials although I would prefer that.
	Back to mammals, lymphocytes and macrophage in mice, any 
suggestions. I was going to look for esterases as a marker for 
macrophage but I'm sure there is something better.
Ian.

Dr. Ian Montgomery,
Microscopy Service Unit,
Graham Kerr Building,
Institute of Biological & Life Sciences,
University of Glasgow,
Glasgow,
G12 8QQ.
Tel: 0141 332 8855 Extn.6644.
e-mail: ian.montgomery@bio.gla.ac.uk



Dr. Ian Montgomery,
Microscopy Service Unit,
Graham Kerr Building,
Institute of Biological & Life Sciences,
University of Glasgow,
Glasgow,
G12 8QQ.
Tel: 0141 332 8855 Extn.6644.
e-mail: ian.montgomery@bio.gla.ac.uk

-- 
Philip Oshel
Supervisor, AMFSC and BBPIC microscopy facilities
Department of Animal Sciences
University of Wisconsin
1675 Observatory Drive
Madison,  WI  53706 - 1284
voice: (608) 263-4162
fax: (608) 262-5157 (dept. fax)