Aniline blue orange g

Wounded mice were euthanized at day 13 after injury. Biopsies from skin wounds were isolated from the back, fixed in formalin and embedded in paraffin. Four-micrometer sections were rehydrated and stained using 0.1% (w/v) Azocarmine G (Sigma, Taufkirchen, Germany) in 1% (v/v) acetic acid for 7 min. Sections were washed with 1% (v/v) acetic acid followed by oxidation in 5% (w/v) wolframato phosphoric acid (Sigma) for 20 minutes and rinsed in water. Sections were then counterstained using 1.5% (w/v) Aniline Blue/Orange G (Sigma) for 4 minutes, washed in water, dehydrated and mounted using Entellan^® water-free mounting medium (Merck, Darmstadt, Germany).

Wound biopsies were isolated and fixed overnight at 4 ℃ in a 4.0% (v/v) formaldehyde solution, pH 7.0 (ROTI^® Histofix 4%, Carl Roth, Karlsruhe, Germany) and embedded in paraffin. Three-micron wound sections were rehydrated and stained in 0.1% (w/v) azocarmine G (Sigma) in 1% (v/v) acetic acid for 7 min, washed briefly with 1% (v/v) acetic acid and followed by oxidation in 5% (w/v) phosphotungstic acid (Sigma) for 20 min and then briefly washed in water. The sections were then counterstained in 1.5% (w/v) aniline blue/orange G (Sigma) for 4 min, briefly washed in water, dehydrated, and mounted in Entellan (Merck, Darmstadt, Germany).