Correlative Microscopy Workflows to Link Functional Spine Imaging and Volume EM

Tuesday, October 5 | 2:00 pm EDT

Dr. Naomi Kamasawa of the Max Planck Florida Institute for Neuroscience will present correlative light and electron microscopy (CLEM) workflows including practical tips and personal experiences. Advanced registration is required.

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Correlative light and electron microscopy (CLEM) is a powerful method to investigate the structure-function relationship in biology. Activities of cells can be recorded using advanced light microscopy (LM), while electron microscopy (EM) provides detailed ultrastructural information in a fixed state. We established two different CLEM workflows to visualize the ultrastructural basis of neuronal functions with volume EM. First, we captured functional properties of the dendritic spines of neurons with live 2-photon Ca2+ imaging, and targeted the same cell, dendrites, and spines in serial block-face scanning EM (SEM) to analyze the structural characteristics of synapses and their surroundings. Second, we induced morphological changes in spines using glutamate uncaging, applied an immuno-EM protocol after fixing the tissues, and performed serial-section array tomography SEM to identify the exact same spine among other unstimulated spines. This webinar covers several practical tips for finding “a needle in a haystack,” including the optimization of sample preparation protocols, selection of detectors for SEM imaging, and application of the focal charge compensation device. Ideas will also be provided on how to choose the best workflow depending on your research goals. In addition, an advanced cryo-CLEM workflow for capturing biological phenomena that require a better temporal resolution is introduced. We used the ZEISS cryo-Airyscan technique for imaging cryo-fixed cultured brain slices followed by array tomography SEM.

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About Naomi Kamasawa
Dr. Naomi Kamasawa has a wide breadth of experience in electron microscopy (EM) stemming from her unique career handling different kinds of biological specimens including bacteria, yeasts, plants, cultured cells, and tissues of invertebrates and vertebrates. As an electron microscopist, she is very enthusiastic about visualizing high-resolution morphology in order to understand physiological function. When she joined Prof. John Rash’s laboratory at Colorado State University in 2003, she became involved in neuroscience research and gained expertise in a very rare EM technique called freeze-fracture replica immunogold labeling, which only a few laboratories in the world are able to perform. She further developed her expertise in visualizing synaptic structure in Prof. Ryuichi Shigemoto’s laboratory at the National institute for Physiological Sciences in Japan, then joined Max Planck Florida Institute for Neuroscience in 2011 as the Head of the EM Core facility. Currently, as the Head of the Imaging Center, she is working with her team to develop state-of-the-art visualization techniques for her collaborative projects, which have been published in many notable journals, including Cell, Nature, Neuron, and eLife.

About the Max Planck Florida Institute for Neuroscience
The Max Planck Florida Institute for Neuroscience (MPFI), a not-for-profit research organization, is part of the world-renowned Max Planck Society, Germany’s most successful research organization with over 80 institutes worldwide. Since its establishment in 1948, 20 Nobel laureates have emerged from the ranks of its scientists, including two announced in 2020. As its first U.S. institution, MPFI provides exceptional neuroscientists from around the world with the resources and technology to answer fundamental questions about brain development and function. MPFI researchers employ a curiosity-driven approach to science to develop new technologies that make groundbreaking scientific discoveries possible. For more information, visit http://www.mpfi.org/.