An improved immunofluorescence technique for the histological examination of blood vessel tissue

Autofluorescence of elastic fibres in blood vessel samples is a common interference with the specific fluorescence of FITC-conjugated antibodies. Counterstaining with eriochrome black T changed the yellow-green colour of elastic fibres to dark red, thus turning a disturbing feature into a useful reference background. A second counterstain, p-phenylenediamine, visualized cell nuclei as an amber colour. To demonstrate the improvement of this staining technique, cryosections from blood vessel samples, derived from control veins, arteries and experimental aneurysms of different ages (15 to 99 month old) in 5 sheep, were stained with antibodies against procollagen III, collagen type IV, laminin, and nidogen. The specific distribution of these connective tissue components could now be related to the location of the elastic fibres and the cells (cell nuclei).