Theta rotation and serial registration of light microscopical images using a novel camera rotating device.

An electromechanical video camera coupler was developed to rotate a light microscope field of view (FOV) in real time without the need to physically rotate the stage or specimen. The device, referred to as the Camera Thetarotator, rotated microscopical views 240 degrees to assist microscopists to orient specimens within the FOV prior to image capture. The Camera Thetarotator eliminated the effort and artifacts created when rotating photomicrographs using conventional graphics software. The Camera Thetarotator could also be used to semimanually register a dataset of histological sections for three-dimensional (3D) reconstruction by superimposing the transparent, real-time FOV to the previously captured section in the series. When compared to Fourier-based software registration, alignment of serial sections using the Camera Thetarotator was more exact, resulting in more accurate 3D reconstructions with no computer-generated null space. When software-based registration was performed after prealigning sections with the Camera Thetarotator, registration was further enhanced. The Camera Thetarotator expanded microscopical viewing and digital photomicrography and provided a novel, accurate registration method for 3D reconstruction. The Camera Thetarotator would also be useful for performing automated microscopical functions necessary for telemicroscopy, high-throughput image acquisition and analysis, and other light microscopy applications.