Abstract

Ultramicroscopy-UM allows 3D-vizualization of biological specimens with μm-resolution. The spatial intensity distribution and thickness of the light sheet have the foremost impact on the quality of image in UM. In this paper, we present different designs for generating light sheet.

The optical unit in standard UM as well as two optimized designs for generating light sheet are reviewed and discussed. The optical characteristics of their light sheet, length and diameter of the line of focus and spatial intensity distribution along the light sheet, are compared. The experimental results demonstrate a marked improvement in reconstructions of representative biological specimens in the third design utilizing aspheric elements. Furthermore, we present our latest achievements in the clearing process of the specimens as well as detection signals by microscope’s objectives that improve further the image quality.

© 2015 SPIE