Periodic Reporting for period 1 - SOADOPP (Supramolecular organization and dynamics of presynaptic proteins)
Reporting period: 2023-09-01 to 2025-08-31
The human brain operates through trillions of connections, known as synapses. Neuronal cells communicate via these connections by releasing chemical messengers in a process that takes milliseconds. At the heart of this process is a sophisticated machinery of proteins that must assemble and act with nanoscale precision. However, the precise three-dimensional arrangement and rapid dynamics of these proteins have remained elusive for decades, hindering our understanding of healthy brain function and the origins of neurological disorders. The SOADOPP project aimed to overcome this fundamental barrier by developing and applying revolutionary super-resolution microscopy techniques. The project's overarching goal was to visualize the 3D supramolecular structures of the presynaptic protein machinery during neurotransmitter release with nanometer resolution. The project combined cutting-edge methods such as Metal-Induced Energy Transfer (MIET), DNA-PAINT and MINFLUX to provide an unprecedented view of this vital biological process.
Throughout the fellowship, the project's central focus was to understand the intricate protein machinery that drives synaptic vesicle release in neurons at the nanoscale. Achieving this ambitious goal required major methodological breakthroughs, which led to significant achievements that have been published in high-impact journals.
A critical first step was to develop a more powerful imaging technique capable of resolving densely packed synaptic structures. This led to the creation of a novel method called fluorescence-lifetime image scanning microscopy SMLM (FL-iSMLM), published in Nature Photonics. This powerful new approach doubles the achievable resolution of standard SMLM and was immediately applied to visualize synaptic scaffold proteins with exceptional detail, demonstrating its power for answering fundamental questions in neurobiology.
Concurrently, to ensure that these unprecedented images of neuronal structures were accurate and reliable, it was essential to develop a new validation tool. This resulted in the introduction of the bacteriophage T4 as a versatile 3D "Bio-NanoRuler," published in Advanced Materials. This work established a simple and robust biological standard, which is critical for validating the microscope's resolution and ensuring that measurements of synaptic protein arrangements are accurate and reliable.
A critical first step was to develop a more powerful imaging technique capable of resolving densely packed synaptic structures. This led to the creation of a novel method called fluorescence-lifetime image scanning microscopy SMLM (FL-iSMLM), published in Nature Photonics. This powerful new approach doubles the achievable resolution of standard SMLM and was immediately applied to visualize synaptic scaffold proteins with exceptional detail, demonstrating its power for answering fundamental questions in neurobiology.
Concurrently, to ensure that these unprecedented images of neuronal structures were accurate and reliable, it was essential to develop a new validation tool. This resulted in the introduction of the bacteriophage T4 as a versatile 3D "Bio-NanoRuler," published in Advanced Materials. This work established a simple and robust biological standard, which is critical for validating the microscope's resolution and ensuring that measurements of synaptic protein arrangements are accurate and reliable.
The results of the SOADOPP project represent a significant step forward, equipping the neuroscience community with powerful new tools for visualising the molecular machinery within neurons. The development of FL-iSMLM overcomes a major obstacle in super-resolution microscopy, enabling researchers to double the resolution of traditional SMLM experiments and observe synaptic proteins with far greater detail. To ensure the accuracy of these advanced neural images, the project also introduced the T4 bio-nanoruler, which is an accessible and much-needed standard for calibrating the three-dimensional performance of the microscopes used for these sensitive measurements. Together, these validated technologies offer a clear path to investigating the molecular basis of synaptic function with unparalleled clarity. The broader societal impact is profound: these tools empower researchers to explore the subtle biological processes at the nanoscale that underlie all neuronal activities, helping to address devastating neurological and psychiatric disorders.