Swinburne University Bionanoengineering Group

Swinburne University Bionanoengineering Group

Swinburne University of Technology Bionanoengineering Group Group Leader: Dan Nicolau Postdoctoral Researchers: Kristi Hanson Prashant Sawant Postgraduate Researchers: Luisa Filipponi Peter Livingston Gerardin Solana Codrin Mocanasu Primary Fluorescence-Relevant Research Application of lithographic techniques for micro/nanostructuring of polymer surfaces to: Control of in vitro motility of protein molecular motors Confinement of live cells, esp. fungi Spatial control of protein attachment with retention of bioactivity Actin filaments (green) moving on a patterned myosin surface. BSA (red) was patterned by CP and used as a myosin blocking agent. Actin filaments moving in tracks created by laser ablation of AAPO. Selective adsorption of FITC-avidin on hydrophobic micro-scale patterns created by bilayer lithography (PtBMA DNQ).

Fungal filament growth in confined PDMS microstructures. Progress, Challenges and Approaches Fungi Progress Growth of fungi in microconfined conditions has revealed a number of unusual phenomena: Challenges Collision-induced branching Directional memory Intra-hyphal coordination Live imaging of cytoskeletal dynamics desirable Requires development and optimization of suitable fluorescent probes Approaches Phalloidin conjugated probes

Issues with toxicity and growth disruption GFP tagged proteins Would require extensive experimental work to develop GFP constructs specific to application relatively little work in this area with fungi Molecular Motors ??? Aim Challenges Growing hyphal tips stained with FM4-64, showing apical vesicle cluster, vacuolar membranes, and mitochondria (from Fischer-Parton et al., 2000). Development of nano-devices able to detect single biomolecules based on actin-bound antibody-antigen interactions. Requires ability to image multiple fluorophores with sufficient temporal and spatial resolution High temporal resolution required to detect effects of interactions on motility

Large field of view at high spatial resolution = low temporal resolution Confocal imaging of only one label at sufficient temporal resolution is already problematic made worse by addition of multiple flourophores Also complicated by interference from polymer substrate, even with confocality, due to nm-scale separation between substrate and protein. Approaches ???? Challenges Specific Challenges Common theme is certainly development of better labels for live cell imaging Chrombodies? Nanoparticle probes? Overall Network Benefit primarily from having access to expertise and resources in other areas, not necessarily requiring extensive development E.g., FCS for monitoring actin-bound antibody/antigen interactions

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