"Establishing fast superresolution live cell and tissue imaging with photomanipulation"

Human immune system is critical for active health and for resolving disease, but information is missing. Over recent years it has become widely accepted in the biomedical sciences that the immune response depends on the multiscale relationships of its organs, immune cells, and molecules, but a holistic perspective remains elusive. Microscopy has, for many years, been a source of scientific discovery for the immune response. However, until recently there has been no microscopy technology with the right sensitivity to study the immune response across scales.

The focus of this MRC Mid-range Equipment application sits at the heart of this challenge aiming to establish the ZEISS Elyra 7 SIM 2 with custom-built photomanipuation, which offers fast quantitative superresolution bioimaging of living tissues across scales down to cells and molecules. The special photomanipulation feature provided by this ZEISS Elyra with photomanipulation allows to photoactivate/photouncage/photobleach and thus track specific molecules and/or cells in the surrounding tissue context.

We tested the ZEISS Elyra 7 at multiple occasions and found the instrument to be ideal for the proposed research questions, well engineered, and user friendly in direct comparison with experiments acquired at bespoke SIM-based microscopes. The ZEISS Elyra 7 was particularly impressive in this respect, allowing one to easily mount and locate samples, as well as maintain SIM under very stable conditions suitable for extended single cell and thin 10-100um tissue slice bioimaging. To the best of our knowledge, this will be the only ZEISS Elyra 7 SIM 2 platform with photomanipulation in the UK.

The Zeiss Elyra 7 SIM2 with photomanipulation meets all the requirements demanded by the tissue community for years. The user base for the ZEISS Elyra 7 with photomanipulation stems from a consortium of MRC-funded and Wellcome-funded investigators and will be undertaking research within UKRI-MRC remit, focusing on infections and immunity, global health, neuroscience and mental health, MRC response to COVID, and translational medicine.

A successful MRC Mid-range Equipment application for the ZEISS Elyra 7 would significantly expand the breadth, capacity, and capabilities of bioimaging in Oxford and in the UK, keeping UK's bioimaging abilities at the forefront of international research.

The MRC Mid-range Equipment in Biomedical Research application will be used to purchase the ZEISS Elyra 7 SIM2 with photomanipuation.

This Elyra platform with photomanipulation offers fast quantitative superresolution bioimaging of thin living tissues across scales down to cells and molecules, which has been an unmet need for years and will be the only platform in the UK. This Elyra platform will be the first commercially available platform custom-equipped with four channel optogenetic photomanipulation for simultaneous widefield or lattice structured illumination microscopy-based (L-SIM) imaging with photomanipulation. This system represents a turn-key solution with an easy to navigate ZEISS software leveraging the latest advancements in microscopy to deliver unprecedented spatiotemporal sensitivity coupled with photomanipulation. The Elyra enables double the conventional SIM resolution allowing researchers to discriminate the finest suborganelle structures, down to 60 nm apart in single cells or between cells across whole tissues thanks to the high temporal resolution. The custom-built photomanipulation feature will enable researcher to photoactivate/photouncage/photobleach, image, and track specific molecules across live cells and thin live tissue explants.

The ZEISS Elyra 7 will be housed and technically supported in a dedicated space at the Oxford-ZEISS CoE and WTCHG imaging facilities. The system will be managed by the facilities staff Drs Lagerholm and Bancroft, an Bioimage Analyst, and further supported by the Oxford-ZEISS CoE and WTCHG teams and resources.

The user base for the ZEISS Elyra 7 with photomanipulation stems from a consortium of MRC-funded and Wellcome-funded investigators and will be undertaking research within UKRI-MRC remit, focusing on infections and immunity, global health, neuroscience and mental health, MRC response to COVID, and translational medicine.