3D-Visualization of nerve fiber bundles by ultramicroscopy

Abstract

Abstract Ultramicroscopy has recently been shown to be an outstanding tool for the 3D-visualization of large microscopical structures with μm resolution. In ultramicroscopy, mechanical slicing is replaced by optical sectioning which eliminates drawbacks such as mechanical distortion and misalignment. Simulations of the illumination light sheet show that the optical sectioning quality of an ultramicroscope greatly depends on the choice of an appropriate slit aperture, thereby the distance between specimen chamber and cylinder lens has to be considered. We labeled nerve fibers in mouse embryos by binding primary antibodies to neurofilament-160 in combination with fluorescent Alexa 488 secondary antibodies. After non-blind deconvolution, 3D-reconstructions of even fine nerve fiber branches were possible. These results demonstrate that ultramicroscopy is a valuable tool for developmental studies in embryology.