EVG staining, the full name is Verhoeff’s Van Gieson staining, it can show black elastic fibers and red collagenous fibers. It can be applied in demonstrating atrophy of elastic tissue in emphysema, thinning and loss of elastic finers in arteriosclerosis and other vascular disease. For this staining solution, it’s stained with hematoxylin containing ferric chloride and iodine firstly, then use excessive mordant to interrupt the combination of tissue and mordant to complete differentiation, then use counterstained with acid magenta.
1.Paraffin section deparaffinating to water:Put the sections into Xylene Ⅰ 20min- Xylene Ⅱ 20min- Anhydrous ethanol Ⅰ 5min- Anhydrous ethanol Ⅱ 5min-75% alcohol 5min, then wash by water;
2.EVG staining:mix alcoholic hematoxylin solution, ferric trichloride, and iodine solution (5：2：2) to EVG staining solution. Put sections into EVG staining solution for 30min, wash by running water.
3.Background Differentiate:Differentiate by ferric trichloride, wash by running water, repeat the operation, and control the differentiation degree under microscope, until elastic fiber is purple black and the background is gray and almost colorless
4.VG counterstaining: Mix saturation picric acid solution and acid magenta solution in 9:1 to generate VG staining solution, stain for 1-3 min, wash quickly, three-cylinder anhydrous ethanol dehydrate quickly.
5.Seal the sections:Clean xylene is transparent for 1-5min and use neutral gum seal the sections.
6.Microscope inspection, images collection analysis.
Elastic fibers and nucleus are black, collagenous fibers are red, other tissue components are yellow background.
1.Precipitation may present in 10% ferric trichloride solution during preservation. Shake well before use.
2.Ensure that prepare fresh Verhoeff's and Van Gieson's staining solution, please discard it if not use out.
3.Control the differentiation degree of 2% ferric trichloride solution under microscope.
4.The time of Van Gieson's counterstaining can’t be too long, and the differentiation time of Anhydrous ethanol can’t be too long neither.