Development of Well Plate Measurement Technique To Evaluate Isolated Vascular Physiological Responses And Comparison With Classical Organ Bath

Objective: The aim of this project is to modify and improve the multi-well organ bath (MuWOB) model and compare this method with the results of classical organ bath. Method: 96-well plates are displayed with a lens fitted monochron 10 Mp resolution camera attached to the bottom of the plate. The distance of the camera to the plate is reduced to a smaller area using mirrors. The pixels of the vessels were counted from the downloaded image. This data was used to determine the physiological responses of the vessel. Contraction responses (phenylephrine 10-11-10-6 M cumulative and 68 mM KCl) and relaxation responses (acetylcholine 10-11-10-6 M cumulative and SNP (sodium nitroprusside) 10-11-10-6 M cumulative) were measured as physiological responses of the vessel. Results: In the modified MuWOB system, distortion disappeared and images were obtained from almost all wells. Comparing the images obtained from the camera, the modified MuWOB was able to produce a clear image in almost all of the plate. In the comparison of the degree of protection of endothelium, it was observed that endothelial responses were better preserved in the modified MuWOB system compared to classical organ bath results. Also, there was no significant difference in the use of gassed and non-gassed Krebs solution in the wells. In the modified MuWOB system, the effect of the refractive index was eliminated by providing visualization from the bottom. Conclusion: The MuWOB system, which is an alternative to an organ bath, has advantages over a conventional organ bath. The modified MuWOB system that we developed can also contribute to the practical use and commercialization of MuWOB.