DHPSFU Fiji Plugin

DHPSFU Fiji Plugin implements an analytical unmixing approach with distance pairing for fast and accurate 3D single-molecule localisation microscopy using the double-helix point-spread function.

Key Features:

• DH-PSF Fitting via Unmixing: Uses an analytical unmixing approach with distance pairing for double-helix point-spread function localisation.
• High Jaccard Index: Achieves a Jaccard index of 0.98 on realistic simulated datasets based on experimental data.
• Fast CPU Processing: Processes localisations at 6,800 locs/s on CPU, exceeding SMAP (2,500 locs/s) and EasyDHPSF (63 locs/s).
• Superior Resolution: Achieves 140 nm resolution when imaging the cellular plasma membrane of Jurkat T cells.
• Scripting Support: Provides Matlab and Python scripts for user customisation.

Scientific Applications:

• 3D Single-Molecule Localisation Microscopy: Enables large depth-of-field 3D localisation microscopy using the double-helix point-spread function.
• Cellular Plasma Membrane Imaging: Applied to imaging the cellular plasma membrane of Jurkat T cells with high resolution.

Methodology:

DHPSFU uses an analytical approach for DH-PSF fitting based on unmixing of fitted localisation data using distance pairing.