Platelet and fibrin deposition to arterial cross-sections detected by indirect immunofluorescence.

Cryosections of allograft samples were perfused with heparinized whole blood and deposited platelets and fibrin were visualized with double immunofluorescence as detailed in Materials and Methods. Representative images illustrate the uneven spatial distribution of GpIIb/IIIa (blue) and fibrin (red) staining in the three layers of the perfused arterial cross-sections. For quantitative evaluation platelet and fibrin surface coverage data were collected from 5–10 such images (original size of the presented images 850 μm × 850 μm) to survey the whole size of the perfused cross-section, and the intrinsic green autofluorescence of elastin (Fig 4, here indicated by the white lines) was used as a guide to encircle the regions of interest and evaluate the three vessel wall layers separately. l: lumen; i: intima; m: media; a: adventitia.