[Histonet] Metachromasia of mast cells in epoxy resin sections

[gillian.2.brown <@t> gsk.com]

John,
could you give me your thoughts then on the methods used in
Interleukin-2–Inducible T Cell Kinase Regulates Mast
Cell Degranulation and Acute Allergic Responses
Johan Forssell et al Am J Respir Cell Mol Biol Vol 32. pp 511–520, 2005,

in which they used and cite the methodology of 
Dvorak AM, Tepper RI, Weller PF, Morgan ES, Estrella P, Monahan ER, Galli 
SJ. 
Piecemeal degranulation of mast cells in the inflammatory
eyelid lesions of interleukin-4 transgenic mice: evidence of mast
cell histamine release in vivo by diamine oxidase-gold enzyme-affinity
ultrastructural cytochemistry. Blood 1994;83:3600–3612.
 and
Oliani SM, Lim LH, Christian HC, Pell K, Das AM, Perretti M. Morphological
alteration of peritoneal mast cells and macrophages in the
mouse peritoneal cavity during the early phases of an allergic 
inflammatory
reaction. Cell Biol Int 2001;25:795–803.

Am I on a hiding to nothing?
Regards

Gill


Target Localisation
Target Discovery
ri- CEDD.
GlaxoSmithKline Medicines Research Centre,
Gunnelswood Road,
STEVENAGE,
Hertfordshire.
  SG1 2NY
tel.          +44 (0)1438 764119
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email.    gillian.2.brown <@t> gsk.com

http://ukdiscovery.gsk.com/histopathology/default.htm




"John A. Kiernan" <jkiernan <@t> uwo.ca> 
07-Oct-2005 18:54
 
To
gillian.2.brown <@t> gsk.com
cc

Subject
Re: [Histonet] Metachromasia of mast cells in epoxy resin sections






I find it hard to believe that the blue or red staining of
mast cell granules, in osmicated and plastic-embedded
tissue, would have any phsiological significance.

I've counted great numbers of degranulating mast cells in
paraffin sections of rat skin and respiratory passages,
after fixation in Carnoy, SUSA and NBF. The granules were
metachromatic (toluidine blue pH 4) inside intact mast cells
and after extrusion from the cells. 
-- 
-------------------------------
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
_______________________________
gillian.2.brown <@t> gsk.com wrote:
> 
> Hi Histonetters,
> 
> would any one have experience of counting granules in mouse mast cells 
in
> tissues which have been formaldehyde fixed, decalcified in a formic acid
> based solution, post osmicated and embedded in epoxy resin?  I'm taking 
1
> micron sections and looking to assess the 'degranulation' status of each
> mast cell in terms of how many are still dark blue and how many are pink
> ie based on the metachromatic properties of toluidine blue.  I suppose 
my
> question is will the formic acid have altered the dye binding properties
> so that I'll get erroneous results?  I'm going to experiment with tissue
> that does not normally have this step but any insights now would be most
> welcome.
> 
> Many thanks
> Gill Brown
> 
> GlaxoSmithKline Medicines Research Centre,
> STEVENAGE,
> 
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