super-resolution-imaging-shared-management-protocol-for-confocal

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Super-resolution microscopy (SRM) overcomes the diffraction limit of conventional light microscopy, enabling nanoscale visualization of subcellular ...

a-multicolor-3d-storm-high-resolution-visualization-protocol-for

A Multicolor 3D-STORM High-Resolution Visualization Protocol for Collagen Mineralization in Self-Assembled Recombinant Type I Collagen Fibrils

This protocol describes a multicolor three-dimensional stochastic optical reconstruction microscopy (3D-STORM) method for nanoscale visualization of ...

Zhejiang University School of Medicine

To study trafficking of bulk internalized vesicles such as macropinosome and lysosome in live cells, an efficient and convenient assay was established according to the axon turning assay. By injecting indicator or fluorescent dyes through a micropipette with air pressure into cell cultures to create a stable gradient around the micropipette tip, vesicles were indicated and labeled. With live cell imaging, the whole process was recorded. Without wash-out of fluorescent dyes and transferring, this assay is an effective, fast labeling system for bulk internalized vesicles, and can also be combined with imaging system.

[Improvement and application of a fast labeling system for bulk internalized vesicles].

Super-Resolution Radial Fluctuation (SRRF) enables live-cell super-resolution imaging, but requires careful parameter selection. Here, we quantify the impact of NanoJ-SRRF parameters on microtubule imaging using FWHM and SQUIRREL-based error mapping. Ring radius proved most critical, with values >1.0 degrading resolution and fidelity. Radiality magnification and axes in ring had minimal impact. Advanced parameters revealed pitfalls: "remove positivity constraint" degraded resolution by 43%, while gradient weighting catastrophically reduced fidelity (RSP = 0.204 ± 0.116). Temporal Radiality Average (TRA) outperformed Temporal Radiality Auto-Correlations (TRAC), milimizing artifacts. This study establishes the first evidence-based guidelines for live-cell tubulin imaging: ring radius ≤1.0, TRA mode prioritization, and avoidance of gradient weighting. Integrating FWHM and SQUIRREL offers a robust opitimization framework for cytoskeletal dynamics.

ArticleTotal : 2	Year
Publication title	2026
Publication title	2026

Article	Year
[Improvement and application of a fast labeling system for bulk internalized vesicles]. Sheng li xue bao : [Acta physiologica Sinica]\| PubMed ID: 29926070	2018
Parameter optimization of NanoJ-SRRF for live-cell microtubule imaging. BioTechniques\| PubMed ID: 40693644	2025

Guifeng Xiao

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