Analyzing isolated blood vessel contraction in multi-well plates

Abstract

Organ baths have been successfully used for over a century to study the contractile or relaxation effects of drugs. Indeed, most of our understanding of vascular pharmacology is based on such in vitro studies. However, multiple parallel organ baths that require mechanical transduction consume relatively large amounts of drugs, gases, and buffers, and they take up a considerable bench space. In addition, such experiments have a high demand in terms of cost and animals, and the tissue preparation is labor intensive and slow. For these reasons, organ baths are no longer in the front line of industrial pharmacological research and they have almost disappeared from most academic laboratories. We have developed a very simple system, which can be implemented virtually in any laboratory, for the automatic analyses of rat aorta ring contraction based on optical methods and using multi-well plates. Rat aorta rings (≈0.5 mm wide) were situated in 96-multi-well plates, and the luminal vessel areas were continuously monitored using a USB camera driven by newly developed algorithms. Liquids were handled using multichannel pipettes, although these procedures can be automated for drug screening. The concentration-response curves obtained were similar to those reported in the literature using traditional force transduction techniques on isolated tissues. This system can also be used with other tissue preparations and for simultaneous fluorescence measurements. The new system described here offers a simple, cheap, and reliable alternative to the classic organ bath system.