Seminars at ASCB | EMBO 2018

December 9, 2018 – 12:30 PM - 1:00 PM | Theater 2, Learning Center

Structured Illumination Microscopy (SIM) with Two-Dimensional Illumination Patterns

Rainer Heintzmann, Ronny Förster, Christian Karras
Institute of Physical Chemistry, Friedrich-Schiller-Universität and Leibniz Institute of Photonic Technology, Jena, Germany

3D-SIM is often a preferred mode of minimally invasive imaging subcellular structures at high resolution. However, it sometimes struggles to deal with densely stained thick samples and with fast sample movements due to the requirement to obtain many, typically 15, raw images for a single high-resolution frame.

Here we analyze the performance of different illumination patterns, including linear gratings and two-dimensional patterns generated incoherently (i.e. sequentially or dithered as in lattice light-sheet microscopy) or by coherent superposition (e.g. in SIM or coherent lattice light-sheet microscopy). Imaging results in various modes are compared in terms of the recently introduced noise-normalized optical transfer function OTFnn. Incoherent superposition is found to be inherently inferior to coherent illumination strategies.

We present a detailed theoretical comparison as well as experimental results on living cells.