Pushing the boundaries of super-resolution optical microscopy for molecular imaging in live tissue

光学显微镜方法改变了我们研究生物体内生物过程的方式，使研究人员能够研究细胞如何生长、分化、交流，以及它们是如何改变以支持细胞可塑性的。这些技术提供了非常高的时空精度和非常低的侵入性，这对于在纳米尺度上观察细胞中的分子动力学和相互作用是必不可少的。这一要求对于研究大脑中支持沟通的机制特别相关，这些沟通是由神经元的微小()亚细胞隔间，即突触介导的。利用新颖的光学纳米技术，我们可以开始在纳米尺度上研究分子机制。然而，随着它们的进一步发展，以及生物成像工具和实验范式的不断演变，这些方法在实施过程中会带来几层复杂性。这限制了它们的采用以及它们在活细胞、组织或体内对多颜色、多模式的适应性。*迫切需要开发成像策略，以提高光学纳米显微镜研究复杂生物范例的可及性和可实现的性能。这方面的进展有可能使光学纳米技术真正改变生命科学。*我项目的长期目标是：*1.建立我的团队和实验室，成为将光学纳米技术和机器学习相结合的新型智能超分辨率方法的领先研究小组，以提高我们对突触通信的理解。*2.开发智能纳米技术平台，允许根据生物样本和实验条件动态调整图像采集。它将集成多模式成像能力，以增强我们探测活神经元突触结构和功能的能力。*该计划将围绕五个短期目标进行阐述：*1.基于深度学习的在线特征识别方法的开发*2.智能扫描系统的实施*3.智能纳米技术与高光谱成像的结合，*4.研究活组织中高光谱纳米技术的新标记物*5.自适应光刺激在功能突触成像中的应用*我的跨学科项目是在光学、化学、机器学习和神经科学的交界处。*智能光学纳米显微镜的拟议开发将建立在我们最近集成的全自动显微镜优化平台的基础上。我提出的NSERC研究计划将为新一代智能成像设备做出贡献，这些设备将改变光学纳米技术的适用性和影响。我的项目的学员将在生物光子学、化学、神经科学和人工智能的界面上发展独特的专业知识。

Optical microscopy methods have transformed the way we study biological processes in living organisms, enabling researchers to study how cells grow, differentiate, communicate and how they are altered to support cellular plasticity. Those techniques offer a very high spatio-temporal precision combined with very low invasiveness, which is essential to observe molecular dynamics and interactions in cells at their scale: the nanoscale. This requirement is particularly relevant for studying the mechanisms that support communication in our brain that is mediated by tiny () subcellular compartments of neurons, the synapses. With novel optical nanoscopy methods we can begin to study molecular mechanisms at the nanoscale. However, as they are further developed, along with the continuous evolution of bio-imaging tools and experimental paradigms, those methods come with several layers of complexity in their implementation. This has limited their adoption as well as their adaptability to multi-color, multi-modal in living cells, tissues or in vivo. ***There is a critical need to develop imaging strategies to increase the accessibility and the achievable performance of optical nanoscopy to study complex biological paradigms. Progress in this direction has the potential to make optical nanoscopy truly transformative in life sciences.***The long-term objectives of my programs are:***1. To establish my team and laboratory as a leading research group on novel intelligent super-resolution methods combining optical nanoscopy and machine learning to improve our understanding of synaptic communication.***2. To develop an intelligent nanoscopy platform that will allow to dynamically adapt the image acquisition to the biological sample and experimental conditions. It will integrate multi-modal imaging capabilities to enhance our ability to probe synaptic structures and function in living neurons.***The Program will articulate around five short term objectives:***1. The development of online feature recognition approaches based on deep learning***2. The implementation of an intelligent scanning system***3. The combination of intelligent nanoscopy with hyperspectral imaging, ***4. The investigation of novel markers for hyperspectral nanoscopy in living tissue***5. The application of adaptive photostimulation for functional synaptic imaging***My transdisciplinary program is at the interface of Optics, Chemistry, Machine learning and Neurosciences. ***The proposed development of intelligent optical nanoscopy will build on our recent integration of a fully-automated microscopy optimization platform. My proposed NSERC research program will contribute to a new generation of intelligent imaging devices that will transform the applicability and the impact of optical nanoscopy. The trainees of my Program will develop a unique expertise at the interface of Biophotonics, Chemistry, Neurosciences and Artificial Intelligence.