Re: Fwd: Re: Fixation buffer.

From:Roger Moretz

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
     NaHCO3             0.197g
     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"
> 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"
> ,
> >
> >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:
> >
> 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:

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