A Multi-modal Imaging Platform Using Light-Sheet and Photostable Probes for Quantitative Cell and Tissue-Scale Biology

Imaging dynamic molecular-scale events with high spatial and temporal resolution is central to discovering fundamental principles underlying molecular self-organization in health and disease.

Achieving this goal requires combined advances in imaging technology and fluorescent probe development applicable to a variety of biological questions and easily adaptable in multiple laboratory settings.

We propose a straightforward light- sheet microscopy design allowing for multi-colour volumetric imaging which in addition enables investigation of molecular properties using multi-modal detection, exploiting a)-fluorescence anisotropy, b)-circularly polarized emission, c)-hyperspectral imaging, and d)-time-stamped detection (fluorescence lifetime, fluorescence fluctuation).

This quantitative plug-and- play imaging platform requires the co-development of bright and photostable probes.

Therefore, we will develop a suit of multi-colour, genetically- encoded, photostable probes based on mStayGold and other recently described fluorescent proteins.

In Phase-1, we will perform feasibility and proof-of- principle studies of the prototype light-sheet system with three detection modalities using existing and new FPs.

We will establish it as a robust and user-friendly platform to then engage with a team of collaborators addressing a breadth of biological questions through investigation of model membranes, cultured cells (yeast, mammalian), and animal models (nematode, fly, fish). Overall, we aim to develop imaging technology including optics and probes for quantitative cell biologists everywhere.