Cell signalling
Can fixation fake phase separation?
Open access · cc by · source: Europe PMC
Comparing the same U2OS cells live vs after PFA showed FET-family IDR puncta increase—and can appear where none existed—warning that fixed-cell LLPS can be artefactual.
Study at a glance
- Design
- Animal / in-vitro — Live vs 4% PFA-fixed imaging of FET-family IDR condensates in U2OS cells
- N
- N=3 · Three FET-family IDRs (FUS, EWS, TAF15) compared live vs fixed
- Population
- U2OS cells transiently expressing EGFP-IDR-NLS constructs
- Outcome
- Fixation-induced changes in apparent LLPS puncta metrics
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Key findings
After fixation, puncta increased in number, size, and contrast; homogeneous EWS-IDR nuclei could gain many discrete puncta; all three quantified metrics rose significantly.
Methodology
Imaged EGFP-tagged FUS/EWS/TAF15 IDRs before and after typical 4% PFA fixation and quantified puncta number, roughness, and punctate percentage.
Limitations
Does not mean all fixed condensates are false, but shows fixation alone can change LLPS appearance.
How this study connects
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