Illumination conditions in microsphere-assisted microscopy
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Updated: Jan 30, 2026

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers
Published on: April 9, 2014
Stephane Perrin1, Hongyu Li1, Audrey Leong-Hoi1
1ICube, University of Strasbourg - UMR CNRS 7357, Photonics Instrumentation and Processes, Illkirch, France.
Abstract:
White-light microsphere-assisted microscopy is a full-field and label-free imaging promising technique making it possible to achieve a subdiffraction lateral resolution. However, performance of this technique depends not only on the geometrical parameters but also on the illumination conditions of the optical system. In the present work, experimental measurements and computer simulations have been performed in air in order to determine the influence of the two diaphragm apertures of the Köhler arrangement and the spectral width of the light source on both the depth-of-focus of the microsphere and the optimization of the imaging contrast. Furthermore, the super-resolution phenomenon is demonstrated and the cumulated optical aberrations are shown through the measurement of the optical transfer function for the different arrangements of the illumination part.
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