The combination of fluorescence light microscopy (FLM) and electron microscopy (EM) makes it possible to put molecular identity in its full ultrastructural context. With array tomography (AT) long ribbons of serial ultrathin sections are labelled via immunohistochemistry (IHC). After FLM imaging the same sections are processed for scanning electron microscopy (SEM) and re-analyzed. The resulting images are correlated and superimposed. Due to the serial nature of this approach it is possible to obtain large volumes of correlated multi-channel light and electron microscopic data. One drawback of this method is the large discrepancy of resolution between light and electron microscopy. To alleviate this we advanced AT for two super-resolution light microscopy techniques, Structured Illumination Microscopy (SIM) and direct Stochastic Optical Reconstruction Microscopy (dSTORM). We also devised a method for easy, precise, and unbiased correlation of EM images and super-resolution imaging data using endogenous cellular landmarks and freely available image processing software. Together, these advances make it possible to map even small subcellular structures with high precision and confidence. We applied our super-resolution AT approach in C. elegans to address an important problem in connectomics research: the mapping of gap junctions at connectomes.
To cite this abstract:Sebastian Markert, Sebastian Britz, Sven Proppert, Marietta Lang, Daniel Witvliet, Ben Mulcahy, Markus Sauer, Mei Zhen, Jean-Louis Bessereau, Christian Stigloher; 3D mapping of subcellular structures with super-resolution array tomography. The 16th European Microscopy Congress, Lyon, France. https://emc-proceedings.com/abstract/3d-mapping-of-subcellular-structures-with-super-resolution-array-tomography/. Accessed: July 13, 2020
EMC Abstracts - https://emc-proceedings.com/abstract/3d-mapping-of-subcellular-structures-with-super-resolution-array-tomography/