3D Reconstruction of Fluorescent Beads

Many optical slices were reconstructed to form 100 views at 3.6 degree intervals around a complete circle. See below for details.


The original image stack was made with a spinning disk confocal microscope consisting of a Zeiss Axiovert 100, Atto confocal unit with arc lamp illumination, and Zeiss 63x 1.4NA plan-Apochromat. The smaller beads are 1um in diameter and larger beads are 4um. The mechanical distance between slices was 0.1um but the optical distance seems to be about twice that. A complete circle of 100 reconstructions was made with some transparency. When the stack is viewed at 90 degrees a regular pattern of vertical lines appears in the larger beads. These lines are probably due to fluctuations in the light intensity (arc lamp flickering) during the acquisition.