ArticleBiology open2016
Critical importance of appropriate fixation conditions for faithful imaging of receptor microclusters.
Article in Biology open, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 52 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
52 citing papers in PubMed, 74 citations in OpenAlex.
- Article
- CaThe Journal of physiology · 2026Article
- Quantitative Characterization of Microtubule Ultrastructure Based on Single-Molecule Localization Microscopy.Cytoskeleton (Hoboken, N.J.) · 2026Article
- Super-Resolution Microscopy Reveals Nanoscale Arrangement of PD-L1 Immune Checkpoint.Chemical & biomedical imaging · 2026Article
- Dual-Color Expansion Microscopy of Membrane Proteins Using Bioorthogonal Labeling.Nano letters · 2026Article
- Time to revise: impact of methodology on boar sperm capacitation in vitro via phosphotyrosine patterns.BMC veterinary research · 2025Review
- Structure and unusual binding mechanism of the hyaluronan receptor LYVE-1 mediating leucocyte entry to lymphatics.Nature communications · 2025Article
- Analysis of immune synapses by τau-STED imaging and 3D-quantitative colocalization of lytic granule markers.Methods in cell biology · 2025Article
- Analysis of Fluorescent Proteins for Observing Single Gene Locus in a Live and FixedThe journal of physical chemistry. B · 2024Article
- Membrane topography and the overestimation of protein clustering in single molecule localisation microscopy - identification and correction.Communications biology · 2024Article
- An aldehyde-crosslinking mitochondrial probe for STED imaging in fixed cells.Proceedings of the National Academy of Sciences of the United States of America · 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
- iU-ExM: nanoscopy of organelles and tissues with iterative ultrastructure expansion microscopy.Nature communications · 2023Article
- Visualizing Protein Localizations in Fixed Cells: Caveats and the Underlying Mechanisms.The journal of physical chemistry. B · 2023Review
- Article
- Tetraspanin-5-mediated MHC class I clustering is required for optimal CD8 T cell activation.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Article
- Chemical fixation creates nanoscale clusters on the cell surface by aggregating membrane proteins.Communications biology · 2022Article
- Understanding immune signaling using advanced imaging techniques.Biochemical Society transactions · 2022Review
- Visualizing the native cellular organization by coupling cryofixation with expansion microscopy (Cryo-ExM).Nature methods · 2022Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors at 2 institutions in 1 country.
Funding
Abstract
Receptor clustering is known to trigger signalling events that contribute to critical changes in cellular functions. Faithful imaging of such clusters by means of fluorescence microscopy relies on the application of adequate cell fixation methods prior to immunolabelling in order to avoid artefactual redistribution by the antibodies themselves. Previous work has highlighted the inadequacy of fixation with paraformaldehyde (PFA) alone for efficient immobilisation of membrane-associated molecules, and the advantages of fixation with PFA in combination with glutaraldehyde (GA). Using fluorescence microscopy, we here highlight how inadequate fixation can lead to the formation of artefactual clustering of receptors in lymphatic endothelial cells, focussing on the transmembrane hyaluronan receptors LYVE-1 and CD44, and the homotypic adhesion molecule CD31, each of which displays their native diffuse surface distribution pattern only when visualised with the right fixation techniques, i.e. PFA/GA in combination. Fluorescence recovery after photobleaching (FRAP) confirms that the artefactual receptor clusters are indeed introduced by residual mobility. In contrast, we observed full immobilisation of membrane proteins in cells that were fixed and then subsequently permeabilised, irrespective of whether the fixative was PFA or PFA/GA in combination. Our study underlines the importance of choosing appropriate sample preparation protocols for preserving authentic receptor organisation in advanced fluorescence microscopy.
Indexed as
Identifiers
What OpenQuestion holds
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.