Measurement of meningeal blood vessel diameter in vivo with a plug-in for ImageJ

Abstract

Changes in blood vessel diameter can be measured manually, but this is time-consuming and often impractical. For automatic measurement commercial solutions are available, but the proprietary algorithms and their potential shortcomings are not known to the user. We present an approach with a CCD camera for image acquisition combined with free and open source ImageJ software for offline analysis. A subtraction image allows for the evaluation of the diameter changes throughout the field of view. A full width at half-maximum algorithm plug-in was written to measure the vessel diameter. For a given line across a vessel, the results of five measurements with parallel shifts throughout an image stack are copied to the clipboard. For validation of this method an established in vivo model was used, namely vascular changes in the rat dura mater, reflecting the activity of the afferent neurons. Vasoconstriction of the meningeal arterioles induced by local electrical stimulation of the dura was inhibited by intravenous administration of the adrenoceptor antagonist phentolamine and amplified by the CGRP receptor antagonist olcegepant. The described methods allow the user to quickly evaluate vessel diameter changes in the whole acquired field at any selected position.