RainbowSTORM: an open-source ImageJ plug-in for spectroscopic single-molecule localization microscopy (sSMLM) data...

Area of Science:

• Microscopy
• Spectroscopy
• Biophysics

Background:

• Spectroscopic single-molecule localization microscopy (sSMLM) offers simultaneous spatial and spectral data acquisition for fluorescent molecules.
• Existing open-source software primarily focuses on localization, lacking comprehensive spectroscopic analysis tools.
• The development of specialized software is crucial for advancing multimolecular and functional imaging capabilities using sSMLM.

Purpose of the Study:

• To introduce RainbowSTORM, an open-source ImageJ/FIJI plugin for comprehensive end-to-end spectroscopic analysis of sSMLM data.
• To provide a user-friendly platform for calibrating, previewing, and quantitatively analyzing emission spectra from sSMLM experiments.
• To address the unmet need for accessible software solutions in sSMLM spectroscopic data processing.

Main Methods:

• Development of RainbowSTORM as a Java plugin for ImageJ/FIJI.
• Implementation of functionalities for spectral calibration, preview, and quantitative analysis.
• Testing across different operating systems (Windows, Mac) and ImageJ/FIJI versions.

Main Results:

• RainbowSTORM provides end-to-end spectroscopic analysis and visualization for sSMLM images.
• The plugin supports the analysis of emission spectra acquired with various sSMLM system designs and fluorescent labels.
• Successful testing confirms the plugin's compatibility and functionality.

Conclusions:

• RainbowSTORM fills a critical gap in open-source software for sSMLM data analysis.
• The plugin empowers researchers to perform detailed spectroscopic analysis, enhancing imaging capabilities.
• RainbowSTORM facilitates the advancement of research in fields utilizing sSMLM techniques.