ArticleScientific reports2015
Image artifacts in single molecule localization microscopy: why optimization of sample preparation protocols matters.
Article in Scientific reports, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 99 papers.
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Who cites it
99 citing papers in PubMed, 206 citations in OpenAlex.
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- Quantitative Characterization of Microtubule Ultrastructure Based on Single-Molecule Localization Microscopy.Cytoskeleton (Hoboken, N.J.) · 2026Article
- Identifying and targeting abnormal mitochondrial localization associated with psychosis.bioRxiv : the preprint server for biology · 2026Article
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- Integrating the Contrasting Perspectives Between the Constrained Disorder Principle and Deterministic Optical Nanoscopy: Enhancing Information Extraction from Imaging of Complex Systems.Bioengineering (Basel, Switzerland) · 2026Article
- Advanced High-Resolution and Super-Resolution Microscopy Techniques for Investigating Chromatin Structure and Epigenetics.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Super-resolution microscopy for structural biology.Nature methods · 2025Review
- Quality control maps: Real-time quantitative control of single-molecule localization microscopy data.Biophysical journal · 2025Article
- Integrating bioengineering, super-resolution microscopy and mechanobiology in autophagy research: addendum to the guidelines (4th edition).Autophagy · 2025Article
- Minimally Invasive DNA-Mediated Photostabilization for Extended Single-Molecule and Super-resolution Imaging.bioRxiv : the preprint server for biology · 2025Article
- Heterogeneity of late endosome/lysosomes shown by multiplexed DNA-PAINT imaging.The Journal of cell biology · 2025Article
- Cryogenic single-molecule fluorescence imaging.BMB reports · 2025Review
- Iterative Immunostaining and NEDD Denoising for Improved Signal-To-Noise Ratio in ExM-LSCM.Bio-protocol · 2024Article
- Improved whole-mount immunofluorescence protocol for consistent and robust labeling of adult Drosophila melanogaster adipose tissue.Biology open · 2024Article
- Exceeding the limit for microscopic image translation with a deep learning-based unified framework.PNAS nexus · 2024Article
- High-density volumetric super-resolution microscopy.Nature communications · 2024Article
- SEMORE: SEgmentation and MORphological fingErprinting by machine learning automates super-resolution data analysis.Nature communications · 2024Article
- Direct observation of the effects of chemical fixation in MNT-1 cells: A SE-ADM and Raman study.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- A Comprehensive Approach to Sample Preparation for Electron Microscopy and the Assessment of Mitochondrial Morphology in Tissue and Cultured Cells.Advanced biology · 2023Article
39 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Single molecule localization microscopy (SMLM) techniques allow for sub-diffraction imaging with spatial resolutions better than 10 nm reported. Much has been discussed relating to different variations of SMLM and all-inclusive microscopes can now be purchased, removing the need for in-house software or hardware development. However, little discussion has occurred examining the reliability and quality of the images being produced, as well as the potential for overlooked preparative artifacts. As a result of the up to an order-of-magnitude improvement in spatial resolution, substantially more detail is observed, including changes in distribution and ultrastructure caused by the many steps required to fix, permeabilize, and stain a sample. Here we systematically investigate many of these steps including different fixatives, fixative concentration, permeabilization concentration and timing, antibody concentration, and buffering. We present three well-optimized fixation protocols for staining microtubules, mitochondria and actin in a mammalian cell line and then discuss various artifacts in relation to images obtained from samples prepared using the protocols. The potential for such errors to go undetected in SMLM images and the complications in defining a 'good' image using previous parameters applied to confocal microscopy are also discussed.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.