Designing and constructing an optical projection tomography microscopy: theoretical and practical considerations

摘要

Optical projection tomography (OPT) microscopy is a new technique to produce high-resolution 3D images of both fluorescent and nonfluorescent biological specimens. It can be used to rapidly map the tissue distribution of RNA and protein expression in intact embryos or organ systems. In optical projection tomography, the specimen is rotated through 360 degrees around a single axis while held in position for imaging. Light passing through specimens is focused onto a digital camera by lenses. After the images of all angles are taken, sections of specimen are independently reconstructed using the filtered back projection algorithm. This paper presents general guidelines for designing and constructing an OPT microscopy. These guidelines are based on the difficulties encountered during the process of building the system and cover theoretical concepts, optical imaging system and reconstruction algorithm required to build the system. In addition, some suggestions are presented to improve future designs.