Probing and imaging cellular and molecular event dynamics at the interface using atomic force microscopy

原子力显微镜（AFM）是纳米科学和纳米技术不可或缺的工具，也是为研究人员提供纳米和亚纳米尺度样品的定性和定量数据的关键系统，包括其表面粗糙度、形貌和机械性能。这些能力使AFM成为申请人研究项目的首选技术，其目的是设计表面和开发表面修饰技术，以增强和促进生物材料与各种生物靶标的相互作用能力，并将其应用于再生医学、纳米医学和芯片实验室设备。

Atomic Force Microscopy (AFM) in its multiple configurations is an integral tool of nanoscience and nanotechnology and a crucial system to provide researchers with qualitative and quantitative data, at nano and sub-nano scale on a sample, including its surface roughness, morphology and mechanical properties. These capabilities make the AFM a technique of choice for the applicants' research program where the aim is to design surfaces and develop surface modification techniques that enhance and promote the interaction capabilities of biomaterials with various biological targets, and this, for their application in regenerative medicine and nanomedicine and lab-on-a chip devices. The PI has acquired an AFM 17 years ago, and since, this equipment has supported not only the PI's research program but also those of the collaborators and many external users. In the last 3 years, the group has faced major problems with the microscope, in terms of acquiring consistent results and image analysis, and since last year the system is not operational at all. This is very harmful to our research as our access is extremely limited to out-campus AFM (nil on-campus et nil during pandemic), especially during regular work hours, adversely affecting measurements, most of which require different time points thus very often losing precious samples. It is urgent to claim our AFM to back in order by replacing defective parts and updating software for data acquisition and image analysis as well as upgrading to enable the applicants to use its full potential for their evolving research program. In this proposal, we have provided examples of applicants' research namely Nanostructured biointerfaces, nanoplexes and biomaterials for cell therapy, tissue engineering and monitoring cellular activities'; Investigation of biomechanics of personalized graft using decellularized matrix of porcine aorta as scaffold'; Non-viral nanoformulations for immunoengineering'; Effect of pathological calcification on soft tissue surfaces'; and Mechanobiology of cells within the periosteum', for which the AFM is critical to reach our research objectives. Through this RTI1 proposal, the applicants request funding to repair, calibrate and upgrade the existing non-functional AFM. This approach is more cost-effective than buying a new microscope, since most hardware parts of the equipment are still in good condition. At this stage, it is crucial that the applicants could benefit from an up-to-date AFM to maintain and enhance its innovative research capacity in regenerative medicine, nanomedicine and diagnostic tools. In addition to enhance our research capacity, the availability of the AFM in our laboratory will provide a transdisciplinary training to the group members and students in which problem-based learning and teamwork are scaffolded to help them, first learn to cross a single disciplinary border and then to cross multiple borders, thus better training them for diverse future careers.