Park, Chan E. Cho, Youngbin Cho, In Jung, Hyunsu Kim, Byeongyeon Shin, Jennifer H. Choi, Sungyoung Kwon, Seok-Kyu Hahn, Young Ki Chang, Jae-Byum 2024-01-19T16:03:21Z 2024-01-19T16:03:21Z 2021-09-02 2020-11-24 1936-0851 https://pubs.kist.re.kr/handle/201004/117826 Actin is an essential protein in almost all life forms. It mediates diverse biological functions, ranging from controlling the shape of cells and cell movements to cargo transport and the formation of synaptic connections. Multiple diseases are closely related to the dysfunction of actin or actin-related proteins. Despite the biological importance of actin, super-resolution imaging of it in tissue is still challenging, as it forms very dense networks in almost all cells inside the tissue. In this work, we demonstrate multiplexed super-resolution volumetric imaging of actin in both cultured cells and mouse brain slices via expansion microscopy (ExM). By introducing a simple labeling process, which enables the anchoring of an actin probe, phalloidin, to a swellable hydrogel, the multiplexed ExM imaging of actin filaments was achieved. We first showed that this technique could visualize the nanoscale details of actin filament organizations in cultured cells. Then, we applied this technique to mouse brain slices and visualized diverse actin organizations, such as the parallel actin filaments along the long axis of dendrites and dense actin structures in postsynaptic spines. We examined the postsynaptic spines in the mouse brain and showed that the organizations of actin filaments are highly diverse. This technique, which enables the high-throughput 60 nm resolution imaging of actin filaments and other proteins in cultured cells and thick tissue slices, would be a useful tool to study the organization of actin filaments in diverse biological circumstances and how they change under pathological conditions. English AMER CHEMICAL SOC Super-Resolution Three-Dimensional Imaging of Actin Filaments in Cultured Cells and the Brain via Expansion Microscopy Article 10.1021/acsnano.0c04915 1 ACS NANO, v.14, no.11, pp.14999 - 15010 ACS NANO 14 11 14999 15010 scie scopus 000595533800049 2-s2.0-85096028293 Chemistry, Multidisciplinary Chemistry, Physical Nanoscience & Nanotechnology Materials Science, Multidisciplinary Chemistry Science & Technology - Other Topics Materials Science Article actin filament expansion microscopy super-resolution imaging synapse postsynaptic density