3D Tessellation Imaging

DESCRIPTION provided by applicant Biology is intricately organized at the nanoscale yet its functional elements such as the neuronal networks in the brain often span over distances of centimeters Ideally tools for observing these networks would provide high resolution over large volumes We propose to meet this challenge by developing D Tessellation Imaging DTI a new super resolution microscopy and combining it with the properties of newly published Expansion Microscopy ExM specimen preparations The combined technology offers nm D resolution Potentially this isotropic high resolution can be achieved in a single exposure per plane an order of magnitude faster than other super resolution methods DTI exploits mathematical symmetries to create sparse crystalline interference patterns These D struc tured illumination patterns probe uorescently labeled structures within a transparent specimen Unlike certain other super resolution techniques DTI is compatible with an essentially arbitrary selection of uorescent labels enabling many color uorescent images ExM physically augments the imaging potential of tissue samples It increases the spatial extent of the sample and makes it easier to see structures that were previously too close together or too small to observe At the same time ExM converts the sample into a hydrogel which is almost entirely transparent but with uorescently labeled locations of interest In this way EM provides both a scalable increase in resolution and a means to observe deeper into tissue volumes In this program we will Build and validate a color objective DTI super resolution prototype Develop an image processing pipeline for raw data from the DTI prototype Demonstrate the powerful synergy of DTI with ExM specimens by creating impressive D brain tissue imagery unattainable by any other method The outcome of these efforts will establish a solid foundation for the continued development of a DTI instru ment that will provide a high resolution brain mapping and tissue mapping tool for biological discovery PUBLIC HEALTH RELEVANCE The complex cellular networks of the human brain are poorly understood Much of the function and disfunction of the brain involves both subcellular structures and cellular interactions that extend over large volumes Investiga tions over this range of spatial scales is dif cult for current technologies D Tessellation Imaging combined with Expansion Microscopy form a promising combination with the potential to map the brainandapos s cellular networks and provide new insights for diagnosis and treatments of brain diseases and disorders