3D and fast imaging: Proposal 2: Spinning disc confocal microscope with laser ablation

• Subject Area: Basic Research in Biology and Medicine
• Term: Funded in 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 563423842

We request an integrated imaging system through a pair of proposals for two complementary microscopes. In Proposal 2, this is for a spinning disc confocal microscope with photomanipulation functionality that includes laser ablation. Key research objectives [identical for proposals 1 and 2]: This proposal unites our expertise under the interdisciplinary framework of “development, health, and eco-interaction dynamics”, with new lines of inquiry arising from joint working across our research systems (11 insect species and the frog Xenopus). For Developmental Biology, this encompasses the regulation of morphogenesis and the roles of major signaling pathways, transcription factors, and downstream effector proteins for shaping the germline and embryogenesis. Basic research insights from this work, embedded in a comparative framework, are of relevance in understanding specific classes of birth defect, also in humans, and in understanding developmental processes that underpin insect species’ biodiversity. Aligning with a growing critical mass at the University of Hohenheim, this proposal also addresses Plant-Insect Interactions, focusing on how insect eggs interact with their oviposition environment at molecular and cellular levels. These two major foci also bring together our research topics in Immunity and Health on responses to wounding, infection, and chemical defense interactions. Description of the instrument: We require a spinning disc confocal microscope for rapid, high resolution imaging of large fields of view, accommodating the tissue-scale focus of many of our morphogenesis investigations. This is combined with a need for high precision photomanipulation, particularly laser ablation (as well as photoconversion and FRET). These features will support our research on: (a) responses to DNA damage; (b) recoil velocity assays of tissue tension; (c) tissue- and stage-specific re-epithelialization dynamics after wounding; and (d) wound response behaviors by living leaf tissue explants as well as insect macrophages (hemocytes) and lamellocytes. Many of these lines of enquiry span spatiotemporal scales from the subcellular (cell junction protein complexes and cytoskeletal regulators) to large tissue fields, and from milliseconds to >24 hours. We are also particularly interested in robust statistical analyses of what are likely to be noisy, somewhat stochastic biological responses. Thus, we require a spinning disc configuration for fast acquisition with low phototoxicity, as well as for tiled acquisition across large tissues and high throughout live imaging of multiple biological samples in parallel.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: 3D und schnelle Bildgebung: Antrag 2: Konfokales Spinning-Disk-Mikroskop mit Laserablation
• Instrumentation Group: 5090 Spezialmikroskope
• Applicant Institution: Universität Hohenheim
• Leader: Professorin Dr. Kristen Panfilio