Table 1 Description of the image analysis process

Blood vessels

Anti CD31 (Dako, clone JC70A, 1/50)

-Scanner: Aperio ScanScope XT (Aperio, Vista, CA)

− × 40 magnification

-TIF format

-Individual images

-TIFF format

-HSV colour model

Ret Fs

Gomori

-Individual images

-JPEG format, quality 80

-RGB colour model

Image-Pro Plus (Media Cybernetics, Inc., Silver Spring, MD, USA)

Customised macro: black and dark brown Ret Fs

-Colour adjustments: None

-Segmentation: R 0–181, G 0–178, B 0–172

-Restriction of the shape and size of the objects detected (roundness >3, area >7.5 μm², length >8 μm, width >1.5 μm) to avoid unspecific signal-form cell nuclei

Col I Fs

Masson's trichrome

Customised macro: specifically detect thick collagen type I bundles and distinguish them from other thinner blue-stained collagen

-Colour adjustments: colour hues intensified (brightness: 68, contrast: 81, gamma: 1)

-Segmentation: R 0–17, G 0–90, B 255–255

-Restriction of the shape and size of the objects detected (roundness >2, area >7.5 μm², length >8 μm, width >10 μm).

Elastic fibres

Orcein

GAGs

Alcian blue pH 2.5 Stain Kit (Artisan, Dako)

-Whole-digitised slide (identification of cylinders corresponding to the different samples)

-TIFF format

-RGB colour model

Aperio ImageScope

Positive pixel count:

-Colour adjustments: None

-Segmentation: RGB >221

-Remaining parameters set by default