Imaging and Volumetric Quantitation of Vascular Corrosion Casts with Laser Scanning Confocal Microscopy.

Abstract

Vascular corrosion casts provide faithful replicas of the three-dimensional anatomy of blood vessel system of organs and tissues. In addition, corrosion casts can be used to obtain quantitative information regarding vessel and tissue spaces. However, morphometric measurements of SEM images of corrosion casts is difficult due to severe specimen tilt, parallax and the inability to image 3D casts from all angles. We present evidence that confocal microscopy can be used to collect all of the 3D information of a corrosion cast in a z-series of optical slices. Surface and volumetric parameters of casted and non-casted space are readily retrievable from a stack of optical sections through a corrosion cast.Various tissues and organs were exsanguinated with heparenized saline following cannulization of regional arteries and casts were made by infusing Mercox resin or Mercox diluted with 20% methyl methacrylate monomer. Tissues were macerated with alternating rinses in 5% KOH and hot water.