How to discern mast cells in toluidine blue stain in mouse brain?

05 August 2022 0 9K Report

I am having trouble evaluating the presence of mast cells in mouse brain sections (formalin-fixed tissue and cryostated onto a slide), specifically within the thalamus. Please see the images attached below.

How would you assess mast cells vs debris or contaminants and what would you reason for the red blotches that appeared through the light microscope on two of the images?

TB Stain Protocol:

Wash slide with absolute alcohol *(8 sections per slide; Pipette used to wash slide)

Wash in absolute alcohol

Wash in 90% alcohol

Wash in 70% alcohol

Wash in running water

Stain sections in TB

Wash in distilled water

Dehydrate with 95% ethanol

*Images 1-4: water and ethanol wash after stain

*Images 5,6: no water and ethanol wash after stain

*Image 1,2: Mouse Atlas slide range- 60-65

*Image 3,4: Mouse Atlas slide range- 50-56

*Image 5,6: Mouse Atlas slide range- 85-88

Link to images: https://drive.google.com/drive/folders/1ubw5OoVFGfhHGIgY93v7zbOE3Dcd738o?usp=sharing

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Eutheria
Rodentia
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