The SFB TRR 274 Bioimaging Platform aims to harness the unique imaging expertise and equipment available at the Munich and Göttingen sites to make cutting-edge intravital biosensor imaging, electron microscopy (EM) and nanoscopy available to all SFB TRR 274 members. For this purpose, we will focus, on the one hand, on establishing an integrated microscopy and IT infrastructure that facilitates access to advanced microscopy techniques and the exchange of imaging data across the SFB TRR 274 sites. This infrastructural integration will benefit a large number of the SFB TRR 274 projects from the start of the first funding phase. In parallel, we will develop novel imaging technologies for multi-scale and multi-parameter imaging. This will expand the resolution range and quantitative accuracy with which cellular, subcellular and molecular checkpoint “settings” can be measured in the living nervous system. Our overarching goal is to establish an easily accessible technology portfolio for performing “in vivo cell biology” of CNS recovery. Thus, the imaging infrastructure and tools developed are aimed to advance the SFB TRR 274’s capabilities to measure checkpoint dynamics at baseline and under therapy. Specifically we aim to achieve the following:
Rationale I • Infrastructural integration
Aim ❶ Build a network of inter-connected CRC imaging units for easy access to intravital, ultrastructural and nanoscopic imaging
We are integrating the imaging units at the SFB TRR 274 sites to provide expertise and access to imaging technologies for intravital and highly resolved visualization of cells and molecules in the CNS to all SFB TRR 274 PIs. This network of imaging units serves as a platform to directly support a large number of our projects as detailed below and builds on highly developed local technology units at all three SFB TRR 274 sites. This platform further provides a web-based hub for storage, documentation, access and exchange of imaging data in order to ease collaboration between sites and allow for data integration across projects.
Rationale II • Technology Development
Aim ❷ Develop quantitative in vivo microscopy approaches to dissect the cell biological and metabolic dynamics underlying CNS recovery
We develop in vivo imaging techniques that use emerging biosensors and quantitative imaging modalities (such as ratiometric FRET and FLIM) to measure the intracellular signals and metabolic states that instruct CNS recovery from single cells to large cellular ensembles.
Coordination: M. Kerschensteiner
Aim ❸ Devise correlated light and electron microscopy techniques for CNS recovery
Correlated light- and electron microscopy (CLEM) approaches are used to superimpose quantitative measurements of subcellular dynamics onto corresponding ultrastructural vistas. We aim to establish two workflows, for either correlating bulk molecular information (based on nanobody stainings and APEX) or individual longitudinal in vivo observations (using laser fiducial marks) with volume EM analysis based on automated ultramicrotomy.
Coordination: M. Schifferer
Aim ❹ Establish correlated in vivo nanoscopy and electron microscopy for molecular structure-to-ultrastructure cross-referencing
We offer nanoscopy as an in vivo technique to visualize subcellular structure and molecular processes within recovering CNS tissue. For this we canl correlate in vivo imaging techniques with improved spatial (nanometre range) resolution with EM (correlated nanoscopy and electron microscopy, CNEM). In its full implementation, this set of technology developments enables the seamless transition between in vivo observations, ultrastructural morphology and molecular tissue structure.
Coordination: S. Jakobs
