Re: Fwd: Re: Fixation buffer.
This is probably waaaay toooo late. However, I
finally found my old copy of Grimstone and Skaer's "A
guidebook to microscopical methods" Cambridge
University Press, 1972. They give a formula for sea
water (pp. 105-109) as:
0.55M NaCl 747.8ml
0.56M KCl 16.3ml
0.38M CaCl2 26.8ml
0.37M MgCl2 145.03ml
0.49M Na2SO4 58.21ml
0.55M KBr 1.5ml
Distilled water to 1000ml
Us old guys have these arcane sources around--it's
just a matter of finding them. The book also includes
the g/l for each of the above, as well as a reference
to the original source as Nicol (1960) "The Biology of
Marine Animals" London: Pitman. There's also data
about pH, buffer qualities, etc. If you need a copy,
send me a fax number and I'll fax it over to you.
--- "Dr. Ian Montgomery"
> 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.
> >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"
> > firstname.lastname@example.org
> >Subject: Re: Fixation buffer.
> >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
> >>Dr. Ian Montgomery,
> >>Microscopy Service Unit,
> >>Graham Kerr Building,
> >>Institute of Biological & Life Sciences,
> >>University of Glasgow,
> >>G12 8QQ.
> >>Tel: 0141 332 8855 Extn.6644.
> >>e-mail: email@example.com
> Dr. Ian Montgomery,
> Microscopy Service Unit,
> Graham Kerr Building,
> Institute of Biological & Life Sciences,
> University of Glasgow,
> G12 8QQ.
> Tel: 0141 332 8855 Extn.6644.
> e-mail: firstname.lastname@example.org
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