Vs asw l100 program

In situ reporter fluorescence-stained slides were analyzed on an EVOS^® FL microscope (Thermo Fisher Scientific). Confocal microscopy was performed using the laser scanning microscope Axio Observer Z1 attached to LSM700 (Carl Zeiss, Jena, Deutschland). Light microscopic cell culture pictures were obtained with an EVOS^® XL microscope (Thermo Fisher Scientific). Total slides were scanned automatically in 40 x magnification using the VS-120-L Olympus slide scanner 100-W system and processed using the Olympus VS-ASW-L100 program (Olympus, Shinjuku, Tokio, Japan).

For documentation, a confocal laser-scanning unit (Axio Observer Z1 attached to LSM710, Zeiss, Göttingen, Germany; ×20 dry and ×40 oil immersion objective lenses, with numeric apertures 0.8 and 1.30, respectively; Zeiss) was used. Sections were imaged using the appropriate filter settings for AF488 (495 nm excitation), tdTomato (554 nm excitation), AF647 (650 nm excitation), and DAPI (345 nm excitation). Up to four channels were detected consecutively and merged. For imaging and documentation of whole optic nerve sections, the Slide Scanner (VS-120-L Olympus slide scanner 100-W system) was used, and images were processed using the Olympus VS-ASW-L100 program (Olympus, Shinjuku, Tokyo, Japan).

For histochemistry and immune‐histochemistry, paraffin‐embedded skin samples were cut into 4 μm sections. Haematoxylin and eosin (HE) staining using Mayer's Hemalaun (1.09249.2500, Merck) and Eosin Y (1.15935.0100, Merck) was done in a linear slide stainer (Leica ST4040). For Masson–Goldner trichrome staining, kits were used according to the manufacturer's recommendations (12043, 14604, Morhisto). Automatic scanning of full slides in 40x magnification was done using the VS‐120‐L Olympus slide scanner 100‐W system and processed using the Olympus VS‐ASW‐L100 program. Evaluation of the epidermal thickness and murine vessel quantification was done according to published work (Ebner‐Peking et al., 2021 (link)), both measured in the Olympus VS‐ASM‐L100 program. Per group, four biological and at least three technical replicates were included.