Verhoeff Van Gieson / EVG Stain
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This method is used for identifying elastic fibers in tissues such as skin, aorta, etc. on formalin-fixed, paraffin-embedded sections, and may be used for frozen sections as well. The elastic fibers will be stained blue-black and background will be stained yellow.
Fixation: 10% formalin.
Section: paraffin sections at 5 um.
1) Deparaffinize sections if necessary and hydrate in distilled
2) Place slides in working Elastic Stain Solution for 15 minutes.
3) Rinse in running tap water until no excess stain remains on
4) Dip slides in Differentiating solution 15-20 times and rinse in tap
5) Check slides microscopically for proper differentiation. Repeat
step 4 if required.
6) Rinse slide in running tap water.
7) Place slides in Sodium Thiosulfate Solution for 1 minute.
8) Rinse slide in running tap water.
9) Stain slide using Van Gieson’s Solution for 2-5 minutes.
10) Rinse in two changes of 95% alcohol.
11) Dehydrate in absolute alcohol.
12) Clear and mount in synthetic resin.
Elastic fibers: black to blue-black
Nuclei: blue to black
Other tissue elements ---------- yellow