High-throughput Lattice Light Sheet Microscopy : Imaging Across Scales.

Biomedical researchers frequently rely on live movies to understand the subcellular cause of diseases and to develop any therapeutic interventions. Such recording of movies of live cells and embryos can be highly phototoxic to the sample and result in cell death within a few hours or a day. To overcome this hurdle, researchers have started using lattice light-sheet microscopes (LLSM) that are extremely gentle and extremely fast with their illumination methodology. In the Greater London Region, we currently do not have a lattice light-sheet microscope in any multi-user facility which is capable of long-term and high-resolution imaging for up to a few days or a week. This is a huge technical disadvantage as several groups are resorting to short-term overnight movies, while these could simply be extended into long-term movies without additional investment into personnel or consumables expenses.

We aim to establish a multi-user LLSM for long-term and high-resolution live-cell studies. In an environment where MRC funds research in a variety of model organisms (from bacteria to human cells), this is a timely and uniquely cost-effective solution to enable World-class Biomedical research and training in the UK.

Ten biomedical researchers studying in a variety of MRC research areas (and funded by MRC) aim to extend their ongoing studies by taking advantage of the Zeiss LLS7, a dual camera-equipped high-throughput microscope. First, with an LLS7, the consortium will be able to move away from short-term movies and start recording long-term movies so that the cause and consequence of any subcellular lesion can be precisely documented in 2D and 3D cell cultures. Second, using the LLS7, the researchers will start tracking the evolution of embryos in the early stages of the worm, fly and fish embryos to precisely link cell-cell interactions and cell fate decisions. Third, MRC-funded structural biologists are also planning to perform high-throughput structure-function studies to accelerate the process of evaluating the immediate and long-term impact of mutants and drug treatment conditions in cells. Together, the three objectives will advance ongoing biomedical research in molecular, cell, developmental and structural biology fields. In addition, by collaborating with Artificial Intelligence experts (in the university and industry) the consortium will set the path for new automated data analysis tools in the emerging area of high-throughput single-cell 'image-omics' field.

Considering the positive impact a long-term imaging system would make in ongoing research, QMUL has agreed to fully match fund the equipment. In addition to accelerating ongoing research, the requested LLS7 microscope supports Queen Mary's ambitious effort in developing a multimillion Precision Healthcare Institute. When the single-cell biology hub of the institute is ready, LLS7 users could extend their work to start studying patient cell fate using dynamic markers. This framework, when available, can help revolutionise therapy strategies - a frontiers healthcare area. Being a leader in high-throughput live imaging for biomedical research could help develop this long-term effort in building highly impactful collaboration(s) with the NHS (for example, newborn or future health teams). Specifically, in collaboration with clinicians, the equipment could be used for the following: (a) to design a single-cell diagnostics pipeline to observe the fate of cells recovered from blood, buccal swabs or urine, (b) to build cellular disease models using patient-derived cells presenting disease-oriented variants and (c) to diagnose the impact of such disease-oriented variants using single-cell based phenotypic reversal assays for precision healthcare.

In summary, securing the LLS7 which is currently unavailable in Europe, will strengthen ongoing internationally leading biomedical research, and in the long-term provide a path to building a patient-facing diagnostic strategy.

To enable high-throughput long-term live-cell imaging across scales, in UKRI/MRC-funded projects, we aim to purchase a lattice light-sheet microscopy. The investigators are already part of a virtual consortium (Center for Cell Dynamics) and use a variety of microscopy methods - Super-Resolution, Deconvolution, Confocal or Electron Microscopy - to study the dynamic function and regulation of biomolecules and subcellular structures in cells, tissues, developing embryos and regenerating worm heads. However, live-cell imaging studies have been limited to Confocal or Deconvolution microscopy that is intrinsically phototoxic, disallowing long-term imaging needed to fully understand health and disease states.

A high-throughput imaging system that allows long-term high-resolution imaging - without inducing photobleaching or phototoxicity - will transform UKRI/MRC-funded studies by allowing the continuous tracking of cell fate, tissue development and regeneration in several model systems, including flies, annelids, zebrafish and human cell cultures. In the long term, such high-throughput facilities can help position us on the frontiers of precision healthcare by providing the high-throughput framework for testing the consequence of human genomic variants, stratifying pathogenic outcomes and testing drug efficacy in 2D cultures or 3D organoids.

Lattice Light Sheet Microscopes (LLSM) use low light illumination allowing long-term live imaging. Their high-speed scans allow high throughput and wide field of view without losing resolution. Multi-user LLSMs in London are all for low throughput studies. A new two-camera LLSM doubles image acquisition speeds, allowing high-throughput dual colour studies of subcellular dynamics in cells, colonies and embryos for days. Because the Zeiss LLS7 requires low maintenance support, it is ideal for a multi-user facility and will significantly transform high-throughput, long-term, live-imaging in the Greater London Region.