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Theta Rotation and Serial Registration of Light Microscopical Images Using a Novel Camera Rotating Device

Published online by Cambridge University Press:  17 March 2010

Bradley S. Duerstock ,
John Cirillo  and
Bartek Rajwa
Bradley S. Duerstock*
Affiliation:
Center for Paralysis Research, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907, USA
John Cirillo
Affiliation:
Center for Paralysis Research, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907, USA
Bartek Rajwa
Affiliation:
Purdue UniversityCytometry Laboratories, Bindley Bioscience Center, Purdue University, West Lafayette, IN 47907, USA
*
Corresponding author. E-mail: bsd@purdue.edu
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Abstract

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° 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.

Keywords

light microscopyphotomicrographydigital video cameratheta rotationimage acquisitionserial section registration3D reconstruction
Type
Instrumentation and Software: Development and Applications
Information
Microscopy and Microanalysis , Volume 16 , Issue 3 , June 2010 , pp. 239 - 247
Copyright
Copyright © Microscopy Society of America 2010

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