Confocal Laser Scanning Microscope

• Subject Area: Neurosciences
• Term: Funded in 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 495215303

This application is for a white-light laser confocal microscope to support the researchprogram of the newly appointed Chair of Organismal and Developmental Neurobiology and the division of Neurobiology at Ludwig Maximilian University. This state-of-the-art imaging system will be incorporated into the Centre for Advanced Light Microscopy, replacing two existing confocal systems that are due to retire in 2022. The underlying concept in this application exploits the existing infrastructure within the Centre for Advanced Light Micorscopy to optimise microscope usage, data acquisition and analysis, and to train and support microscope users. The microscope itself will be critical for the future research programs of the applicants, which focuses on cellular and systemic neuroscience. To address questions in neuroscience in a meaningful way, imaging across different scales (from molecules, to cells, and circuits) has become increasingly important. Accordingly we require a versatile microscope with a number of features. Specifically:(1) A tuneable white light laser.(2) A motorised stage and imaging stitching software.(3) A cell culture chamber.(4) Confocal super resolution capacityA tuneable white light laser will enable the applicants to undertake tracing experiments employing a variety of different fluorescent tracers as well as genetic tools such as theConfetti/Brainbow system to label individual neurons. The ability to define specificwavelengths for excitation will likewise be important for future insitu hybidisation experiments that will use a broad variety of probes simultaneously (e.g. opal 520, Cy3, opal 570, Cy5). A motorised stage and stitching software will facilitate the capture of labelled neurons at high resolution, which will be used to generate large quality mosaics facilitating circuit tracing. A cell culture chamber will enable the quantification of microtubule dynamics, an assessment of mitotic progression in progenitors, and calcium imaging in organotypic neuronal cultures. Finally, we will benefit from a microscope that couples the latest developments in deconvolution algorithms with high sensitivity detectors to image biological structures (e.g. centrosomes, dendritic terminals, nodes of Ranvier) at a near super resolution (120nm). Collectively this microscope will form a bedrock for the Neurobiology community at Ludwig Maximilian University, equipping us with the capacity to undertake cutting-edge studies for the next decade.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Laser Scanning Konfokalmikroskop
• Instrumentation Group: 5090 Spezialmikroskope
• Applicant Institution: Ludwig-Maximilians-Universität München
• Leader: Professor David Keays, Ph.D.