In the past decade, research in development and oncology has uncovered an impressive number of relevant signaling pathways. While a qualitative understanding of signals, and their connection to cellular outputs has been established, plasticity and feedback of signaling networks remain obscured. A better understanding of dynamic and locally constrained signaling events driving organ development and disease progression requires access to refined subcellular probe detection. The availability of optogenetic and chemical biology tools provides novel opportunities, but requires dedicated microscopy equiment. For this reason, six projects at the Department of Biomedicine (DBM) and the Biocenter (BC) of the Univerisity of Basel i) localized lipid signaling in disease (M. Wymann, DBM); ii) dynamic subcellular Wnt/b-catenin signaling in epithelial mesenchymal transition (G. Christofori, DBM); iii) DNA dynamics and confined epigenetic plasticity (P. Schär, DBM); iv) real-time monitoring of Sonic Hedgehog and Bone Morphogenetic Protein gradients in limb buds (R. Zeller, DBM); v) ultrastructural analysis of neuronal stem cell control (V. Taylor, DBM); and vi) molecular mechanisms determining the development of vascular networks (M. Affolter, BC), illustrate the need of the requested “subcellular targeting microscopy” equipment. The core of the platform is a highly sensitive microlens-enhanced spinning disk microscope linked to FRAP, ablation, and multiple excitation laser lines, and an integrated TIRF module to monitor plasma membrane events.

Through its integration into the BioOptics core facility at the DBM, the subcellular targeting platform will be accessible to >500 regional researchers at the University of Basel, DBM, FMI, D-BSSE, Fachhochschule, etc. An image storage and analysis pipeline with remote user access capabilitiy is in place, to allow seamless operation and output.

The requested equipment will allow the use of genetically encoded opto-genetic proteins, proteins tagged for reactivity with chemical inducers of dimerisation (CIDs), and the possibility to perform FRAP/TIRF and FRAP/confocal microscopy, and will greatly enhance the possibilities to manipulate and track subcellular localization of target proteins. The insights gained by these experimental approaches will be critical for a better understanding of dynamic biological processes, and will spur the design of innovative therapeutic approaches to counteract resistance mechanisms in oncology.