Fluorescent Microscopy Imaging with DAPI stain to assess the endothelial Integrity in Goat Artery Strip

Abstract

Isolated blood vessel experiments are commonly done to  understand the dynamic functions of vascular smooth muscle, pharmacology of various receptor subtypes and influence of the endothelium on vascular biology. Experiments can be done either with endothelium-intact or with denuded vessel strip, as required. Since vascular biology can be modulated by the endothelium, it is necessary to confirm the presence or absence of endothelium in an endothelium-intact or denuded vessel respectively. To achieve the above strategy, isolated arterial strips from goat (Capra hircus) leg was segregated into a control group with intact-endothelium & other group with endothelium removed. After fixation, paraffin blocks were made with impregnated tissue & tissue sections were made using Leica microtome, which were then stained with DAPI (4',6-diamidino-2-phenylindole), a nuclear stain and images were taken under fluorescent microscope. Results show a layer of nuclei in endothelium-intact vessel and not seen in endothelium-denuded vessel, confirming the presence and absence of endothelium respectively. Hence,fluorescent   microscopy imaging of tissue section with DAPI stain will be a reliable method to assess the endothelial integrity in blood vessel.

Keywords: Fluorescent microscopy,Goat artery,DAPI stain, Endothelium