Nanoscale Characterisation of Biological and Bioinspired Materials using Integrated Fluidic Force - High-Resolution Confocal Microscopy

We propose a new imaging platform that combines ultra-fast confocal imaging with the the nano-fluidic functionality delivered by an integrated Fluidic Force microscope (FluidFM-UFCLSM). The proposed capability opens a new phase of exploration of biological systems by enabling characterisation of localised biochemical and physiological processes. The proposed capability provides new avenues for specific applications such as new antimicrobial agents, functional genetics and the development of sustainable crops. The unique design of FluidFM-UFCLSM enables accommodating an array of complex biological samples to perform quantitative and predictive characterisation of biofilms, tissues, whole plants, small animals, insects, mucosal membranes, food systems and tissue scaffold hydrogels. The unique feature of FluidFM-UFCLSM is it will enable study of the smallest units of biological organisation such as proteins as well as larger objects such as cells, tissues and organs. The use of FluidFM-UFCLSM cuts across many disciplines and delivers benefits to a broad range of research topics in the areas of biofilm formation, plant science, tissue engineering, food science and cell physiology. Some examples of FluidFM-UFCLSM applications are:
1) Elucidate anti-microbial resistance and the localised mechanisms underpinning quorum sensing
1) Probe interaction between immune cells with lung epithelium as one of the key pathways of Covid-19 pathogenies
2) Uncover the secrets of plant development and mechanical signalling to develop new resistant crops
3) Probe the effect of nutrition on gut microbiome and associated health outcomes
4) Explore new plant-mimetic materials for designing new food-compatible films for environmentally sustainable food production
The broader areas of impact will be achieved by supporting emerging areas research that targets the major problems and challenges of food security, improved nutrition, animal and human health, combatting antimicrobial resistance, microbiome research, industrial biotechnology, waste valorisation, sustainable agricultural and synthetic biology.

The proposed capability is enabled by integrating an open architecture Fluidic Force Microscope (FluidFM) with a Confocal Laser Scanning Microscope (CLSM), equipped with an ultra-fast (UF) scanning unit and automated multi-scale (from nano to sub-millimetre) positioning system with a large Z-range. Thus, the proposed capability enables exploration of biological systems using an optically guided fluidic component to enable: a) delivering target chemicals, such as drugs, gene editing enzymes and antibodies to the cell surfaces and tissues; b) utilising FluidFM cantilever as a micropipette to manipulate cells and other particulate objects, such as viral capsids; and c) using FluidFM to perform nanoscale patterning and 3D printing.
Due to a large XYZ range (>100 um for FluidFM and 500 um for CLSM) and 10 mm sample space, the FluidFM-UFCLSM uniquely enables accommodating complex biological systems and challenging samples such as biofilms, whole plants and small animals. These characteristics make the proposed equipment stand out on the global arena.
In addition to ultra-fast scanning, the FluidFM-UFCLSM will also have high-resolution capabilities, and will enable correlative imaging down to < 120 nm resolution, on a single platform.
The proposed equipment will have a motorised micro-positioning stage and automated cross-talking of assimilated software in order to enable high-speed analysis at multiple areas of interest and accelerated testing capabilities. Image analysis of large arrays of image data requires new approaches to data analysis, which will include recently developed techniques in machine learning for the analysis of complex multi-dimensional data. This will allow the routine analysis of high-dimensional data at a speed that can match the data capture, with a minimum requirement on researcher time.
The equipment will be housed in the Containment Level 2 environment to enable accommodating biological samples and Hazard group 2 biological agents.