Fast scanning two photon microscope with confocal unit for intravital and reflectance imaging

• Subject Area: Neurosciences
• Term: Funded in 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 467868227

The present proposal concerns a state of the art two photon microscope capable to record simultaneous multi-chromatic functional signals, such as calcium events, and perform high resolution subcellular morphological recordings on large fields of view in freely behaving animals.The requested equipment is central to the successful establishment of Dr. Nicolas Snaidero’s research group at the Hertie Institute for Clinical Brain research (HIH) and will in addition, provide technical solutions for many well establishes groups at the HIH and at the Center for Integrated Neuroscience (CIN) as part of the virtual imaging facility between these institutes. Indeed, currently the HIH and CIN have several two photon microscopes that are already heavily used and not suitable for the described projects due to major technical limitations. Indeed the innovative projects described in this application make this instrument absolutely necessary for the applicants and the entire institution.The Major scientific targets of the research groups listed in this proposal focus on the functional and morphological interaction between glial, epithelial, immune and neuronal cells in the brain and spinal cord but also in developing organoids and organs such as lung and liver. Advanced two photon imaging will be combined with complex models, behavioral studies and translational approaches to investigate axonal and glial pathologies as well as migration of immune cells in inflamed milieu, plaque dynamics and immune modulation in neurodegenerative disease models, neurotransmitter and hyperactivity on connectivity and brain development.To achieve these objectives a multi photon microscope equipped with two Ti:Sapphire lasers, one of which with an extended IR range up to 1300nm combined with fast resonant tandem scanner and high performance non descanned external detectors is required. The versatile nature of the requested system with adjustable excitation/detection allows optimal imaging of a wide variety of fluorophores and the adaptive stage is compatible with large behavioral setups. Furthermore, the integration of confocal components allows concomitant 2P and Spectral Confocal Reflectance Microscopy to combine label free imaging of myelin sheath with intravital imaging. Finally, the possibility for the system to be upgraded in order to perform 2P intravital Fluorescence Life time Imaging offers the unique opportunity to broaden dynamic applications especially towards metabolic state investigations and subcellular molecular changes.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Zwei-Photonen Mikroskop mit konfokaler Einheit für Intravital- und Reflexionsbildgebung
• Instrumentation Group: 5090 Spezialmikroskope
• Applicant Institution: Eberhard Karls Universität Tübingen
• Leader: Nicolas Snaidero, Ph.D.