ArticleeLife2021
A robust method for particulate detection of a genetic tag for 3D electron microscopy.
Article in eLife, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 29 citations in OpenAlex.
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- Ribosome-binding protein 1 maintains peroxisome biogenesis.Journal of cell science · 2025Article
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- Single molecule visualization of tropomyosin isoform organization in the mammalian actin cytoskeleton.Cytoskeleton (Hoboken, N.J.) · 2025Article
- Quantitative Fluorescence Analysis Reveals Dendrite-Specific Thalamocortical Plasticity in L5 Pyramidal Neurons during Learning.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2023Article
- A Validated Set of Ascorbate Peroxidase-Based Organelle Markers for Electron Microscopy of Saccharomyces cerevisiae.mSphere · 2022Article
- Volume electron microscopy.Nature reviews. Methods primers · 2022Article
- Article
- Finding Needles in a Haystack with Light: Resolving the Microcircuitry of the Brain with Fluorescence Microscopy.Molecules and cells · 2022Review
- Microscopic Visualization of Cell-Cell Adhesion Complexes at Micro and Nanoscale.Frontiers in cell and developmental biology · 2022Review
- Cavin4 interacts with Bin1 to promote T-tubule formation and stability in developing skeletal muscle.The Journal of cell biology · 2021Article
- GFP-Tagged Protein Detection by Electron Microscopy Using a GBP-APEX Tool inFrontiers in cell and developmental biology · 2021Article
- Niwaki Instead of Random Forests: Targeted Serial Sectioning Scanning Electron Microscopy With Reimaging Capabilities for Exploring Central Nervous System Cell Biology and Pathology.Frontiers in neuroanatomy · 2021Review
Corrections and comments
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Authors and funding
20 authors at 6 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Genetic tags allow rapid localization of tagged proteins in cells and tissues. APEX, an ascorbate peroxidase, has proven to be one of the most versatile and robust genetic tags for ultrastructural localization by electron microscopy (EM). Here, we describe a simple method, APEX-Gold, which converts the diffuse oxidized diaminobenzidine reaction product of APEX into a silver/gold particle akin to that used for immunogold labelling. The method increases the signal-to-noise ratio for EM detection, providing unambiguous detection of the tagged protein, and creates a readily quantifiable particulate signal. We demonstrate the wide applicability of this method for detection of membrane proteins, cytoplasmic proteins, and cytoskeletal proteins. The method can be combined with different EM techniques including fast freezing and freeze substitution, focussed ion beam scanning EM, and electron tomography. Quantitation of expressed APEX-fusion proteins is achievable using membrane vesicles generated by a cell-free expression system. These membrane vesicles possess a defined quantum of signal, which can act as an internal standard for determination of the absolute density of expressed APEX-fusion proteins. Detection of fusion proteins expressed at low levels in cells from CRISPR-edited mice demonstrates the high sensitivity of the APEX-Gold method.
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