ArticleBiophysics reports2017
Determining the target protein localization in 3D using the combination of FIB-SEM and APEX2.
Article in Biophysics reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 15 citations in OpenAlex.
- The Matrix of Mitochondrial Imaging: Exploring Spatial Dimensions.Biomolecules · 2025Review
- DeepContact: High-throughput quantification of membrane contact sites based on electron microscopy imaging.The Journal of cell biology · 2022Article
- Cryogenic superresolution correlative light and electron microscopy on the frontier of subcellular imaging.Biophysical reviews · 2021Review
- Assaying three-dimensional cellular architecture using X-ray tomographic and correlated imaging approaches.The Journal of biological chemistry · 2020Review
- Article
- Gold-substituted Silver-intensified Peroxidase Immunolabeling for FIB-SEM Imaging.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2019Article
- Article
- Ultrastructural localisation of protein interactions using conditionally stable nanobodies.PLoS biology · 2018Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Determining the cellular localization of proteins of interest at nanometer resolution is necessary for elucidating their functions. Besides super-resolution fluorescence microscopy, conventional electron microscopy (EM) combined with immunolabeling or clonable EM tags provides a unique approach to correlate protein localization information and cellular ultrastructural information. However, there are still rare cases of such correlation in three-dimensional (3D) spaces. Here, we developed an approach by combining the focus ion beam scanning electron microscopy (FIB-SEM) and a promising clonable EM tag APEX2 (an enhanced ascorbate peroxidase 2) to determine the target protein localization within 3D cellular ultrastructural context. We further utilized this approach to study the 3D localization of mitochondrial dynamics-related proteins (MiD49/51, Mff, Fis1, and Mfn2) in the cells where the target proteins were overexpressed. We found that all the target proteins were located at the surface of the mitochondrial outer membrane accompanying with mitochondrial clusters. Mid49/51, Mff, and hFis1 spread widely around the mitochondrial surface while Mfn2 only exists at the contact sites.
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