Confocal Laser Scanning Microscope with Spinning Disk Technology

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 522417173

OICE is a centre of excellence (central institute) of FAU and represents the centralised platform for high and ultra-high resolution imaging using light-based technologies for, among other things, living cells and tissues, organoids, intravital imaging and imaging of biological materials and membranes. OICE combines a Core Facility Unit (CFU), Exploratory Research Unit (ERU) and Educational Training Unit (ETU). The interaction of physics/engineering on the development side, with researchers from natural sciences, medicine and material sciences on the application side, and the well-founded training strengthens Erlangen as a research location and has established OICE nationally and also internationally as a recognised institute for light microscopy. The novel combination of real-time super resolution with spinning disc laser scanning allows for the first time long-term measurements of living three-dimensional samples with improved lateral resolution and, in combination with improved silicone immersion objectives, higher penetration depths with lower artefacts of > 100 µm. However, the speed and significantly lower illumination intensity of an SD-LSM is retained. Thus, this Super Resolution SD-LSM for long-term live cell observation replaces the previous SD-LSM (acquired in 2013) and complements the existing devices such as the multiphoton (intravital imaging) LSM and light sheet microscopy (fixed samples). At the OICE, there is still a high demand for spinning disc technology, especially considering the technological advancements in resolution, penetration depth and better camera detectors. The majority of the > 30 research groups/year that successfully use this technology use organoid models, live cell cultures or tissues to perform long-term spatio-temporal multichannel imaging (from several hours to whole days). The highest possible lateral (super) resolution for dynamic intracellular or membrane-associated processes is essential. The high thickness of the samples (organoids / tissue) requires a high penetration depth with minimised scattering to enable meaningful three-dimensional reconstruction and measurements, also over time. A large optical field of view that is as homogeneously illuminated as possible also minimises tile imaging with downstream stitching, which is associated with losses. In the medium term, this device should also relieve and replace the classic SD-LSM, which has now been in continuous operation for ten years (> 2,600 hours/year).

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Konfokales Laser Scanning Mikroskop mit Spinning Disk Technologie
• Instrumentation Group: 5090 Spezialmikroskope
• Applicant Institution: Friedrich-Alexander-Universität Erlangen-Nürnberg
• Leader: Dr. Ralf Palmisano