Vascular fluorescence casting and imaging cryomicrotomy for computerized three-dimensional renal arterial reconstruction

OBJECTIVES To assess the combined use of a casting technique, cryomicrotomy imaging, and three-dimensional (3D) computer analysis as a method for visualizing and reconstructing the arterial vascular tree in a porcine renal model. MATERIAL AND METHODS The arterial branches of two porcine kidneys were filled with a fluorescent cast, after which they were cut into slices of 50 mu m in an imaging cryomicrotome. From each section, digital images of the cutting plane of the sample were taken and stored in the computer, after which stacks of images were rendered in 3D. RESULTS A 3D computerized reconstruction of the arterial vascular tree was constructed and showed the complete arterial anatomy up to arterioles of 50 mu m. CONCLUSION With visualization by fluorescence imaging cryomicrotomy, the anatomical and 3D reconstruction of the renal arterial blood supply in a pig kidney is possible up to a resolution of 50 mu

Porcine coronary collateral formation in the absence of a pressure gradient remote of the ischemic border zone

In the current paradigm on coronary collateral development, it is assumed that these vessels develop consequentially from increased fluid shear stress (FSS) through preexisting collateral arteries. The increased FSS follows from an increase in pressure gradient between the region at risk and well-perfused surroundings. The objective of this study was to test the hypothesis that, in the heart, collateral connections can form in the absence of an increased FFS and consequentially at any depth and region within the ventricular wall. In Yorkshire pigs, gradual left circumflex coronary artery occlusion was obtained over 6 wk by an ameroid constrictor, whereas the control group underwent a sham operation. Hearts were harvested and subsequently processed in an imaging cryomicrotome, resulting in 40-μm voxel resolution three-dimensional reconstructions of the intramural vascular vessels. Dedicated software segmented the intramural vessels and all continuous vascular pathways containing a collateral connection. In the ameroid group, 192 collaterals, 22-1,049 μm in diameter, were detected with 62% within the subendocardium. Sixty percent of collaterals bridged from the left anterior descending artery to left circumflex coronary artery. A novel result is that 25% (n = 48) of smaller-radius collaterals (P = 0.047) connected with both origin and terminus in the nontarget area where perfusion was assumed uncompromised. In the porcine heart, collateral vessels develop not only in ischemic border zones with increased FSS but also away from such border zones where increased FSS is unlikely. The majority of collaterals were located at the subendocardium, corresponding to the region with highest prevalence for ischemi