Automated Camera Lucida Method with Colored Images through Integration of Hardware and Software in Microscopic Zooming.

Abstract

Automating the camera Lucida method which is a standard way for focusing microscopic images is a very challenging study for many scientists. Hence, actually combining hardware and software to automate microscopic imaging systems is one of the most important issues in the field of medicine as well. This idea reduces scanning time and increases the accuracy of user's results in this field. Closed-loop control system has been designed and implemented in the hardware part to move the stage in predefined limits of 15°. This system produces 50 consecutive images from parasites at the mentioned spatial distances in two directions of the z-axis. Then, by introducing our proposed relational software with combining images, a high-contrast image can be presented. This colored image is focused on many subparts of the sample even with different ruggedness. After implementing the closed-loop controller, stages movement was repeated eight times with an average step displacement of 20 μm which were measured in two directions of the z-axis by a digital micrometer. On average, the movement's error was 1 μm. In software, the edge intensity energy index has been calculated for image quality evaluation. The standard camera Lucida method has been simulated with acceptable results based on experts' opinions and also mean squared error parameters. Mechanical movement in stage has an accuracy of about 95% which will meet the expectations of laboratory user. Although output-focused colored images from our combining software can be replaced by the traditional fully accepted Camera Lucida method.