http://www.nature.com/jcbfm/journal/v30/n4/fig_tab/jcbfm2009282f2.html

They seem to have done it.

Another approach is to use a membrane marker for total mast cells (eg IgE Fc, c-Kit) and to compare MC numbers identified by immunohistochemistry with mast cells detected by histochemical staining. This allows identification of even those MC that are fully degranulated. It depends on whether you want to examine acute MC degranulation or more chronic degranulation. With the appropriate fixative, both staining procedures can be done on the same section - I used Carnoy fixation, Alcian Blue staining and detection of IgE in rat tissues and Carnoy/Alcian Blue/c-Kit in human tissues. European journal of immunology. 09/1976; 6(8):537-45. Australian Journal of Experimental Biology and Medical Science 07/1987; 65 ( Pt 3):241-50. DOI:10.1038/icb.1987.27

In general I have to remind that detection of MCs by fluorescent avidin (Jin et al, Stroke 2009; 40: 3107-3112) has the advantage of being nondestructive and can be used in conjunction with other types of analysis, including conventional microscopy to appreciate for example the extent of inflammatory cell infiltration.

Hi everybody who answered

I presented a part of my method here. may you please see is there any mistake in my method:

Under general anesthesia, 5ml normal saline and 5 ml formalin 4% perfused transcardially to wash the vessels from blood and to fix the brain tissue, respectively. The brain extracted and placed in formalin 4% at least for 3 days. Coronal sections of the brain in mid hippocampus and brain stem were stained with Toluidine blue and observed for number and state (degranulation) of mast cells

Hi again

I attached a picture of my study here. please look if you can see any mast cells

Picture 2

Picture 3

Hi, probably there is one mast cell on the right of picture 2. I'm not sure because the magnification is low. How long it takes to stain with Toluidin Blue? What is the buffer that you use? What is the % of toluidin blue in the buffer?. Regards

http://www.ncbi.nlm.nih.gov/pubmed/2578142

http://www.ncbi.nlm.nih.gov/pubmed/16219422

As showed in the paper above, you can use conjugated-avidin to stain brain mast cells.