A multi-modality imaging system for super-resolution (SIM/STED/STORM) applications in space and time

The microscopy unit at the Life Sciences & Engineering (LS&E) infrastructure center applies tools from various fields ranging from optical engineering through signal processing to biology processes. A major effort of the center revolves around super-resolution (SR) microscopy and related applications.

We have constructed a multimodality SR-microscopy system that can interchange between the different SR methods on the same sample. The system was designed as a multiuser microscope that achieves multi-modality and multi-resolution imaging both in space and in time. The multimodal SR-system is based on Elyra7 (Zeiss) microscope that supplies: Structured Illumination Microscopy (SIM²) a ‘real-time’ imaging method for dynamic samples, and single molecule localization modalities PALM\STORM, that can supply a ‘ground truth’ measurement with high precision of localization. In addition, this system includes STEDYCON (Abberior) system for STED imaging, or for laser manipulation/activation combined with the SIM² modality for fast live super-resolution imaging.

We demonstrate that a multi-modality microscope can be used for laser manipulation applications combined with spatial and temporal super-resolution imaging. One example was cells treated with a short-labeled peptide that were imaged using SIM² and STED setups. The images were co-registered in the nanometers scale with IMARIS software. Then, the STED setup was applied to bleach a defined region of interest. Immediately after, SIM² was used to observe live cell dynamics at 40fps. Our combined SR-microscope allowed exploiting the advantage of SIM technique (gentle and fast) for live-cell imaging with the ability of a laser-scanning microscope, provided by STED for laser manipulation applications.