Mast cell staining
From: | "Timothy Martin" <MartinTJ@Cardiff.ac.uk> |
Could anyone give me some information to help clear up and
further understand a rather important bone of contention regarding
staining properties and characteristics of the two different types of
mast cells found in guinea-pig lungs and trachea. I have recently
had my PhD oral exam in Cardiff at the Welsh School of Pharmacy and
the external examiner was took exception to the staining protocol
(Find theprotocol I used below) that I used to stain dispersed
guinea-pig lung and racheal mast cells
it was first stated that the aclian blue and safranin should not be
added at the same time but stained firstly with aclcian blue and then
counterstained with safranin. Secondly it was claimed that guinea-pig
connective tissue mast cells do not stain red with safranin O anyway,
and thirdly it was claimed that fixing the cells was essential for
the staining of these cells that without it the cells would not
stain. He was therefore doubting my interpretation of my results
which were that guinea-pig pig connective tissue mast cells stain red
with safranine O whislt mucosal mast cells stain blue with alcian
blue.
Could you therefore give me any comments on the above, imparticular
the importance of fixing cells on the staining properties of the
alcian blue safranin combined stain on guinea-pig airway mast cells,
and your opinion on whether you think the above protocol was suitable
for the staining of guinea-pig mast cells. Also could you give me any
further information regarding guinea-pig mast cells from the airways
With kind regards
Timothy Martin
Department of Pharmacology
Welsh School of Pharmacy
Cathays Park
Cardiff
UK
CF10 3XF
Email martintj@cardiff.ac.uk
Protocol
Guinea -pig lungs and trachea were first isolated and digested using
collagenase and BSA. After filtering and spinning down the cells,
they
were not fixed and were stained in a solution which contained 0.36%
Alcian blue and 0.2%
Safranine O. This stain was made up in acetate buffer.
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