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

Janel L Davis1, Brian Soetikno1, Ki-Hee Song1

  • 1Department of Biomedical Engineering.

Bioinformatics (Oxford, England)
|July 15, 2020
PubMed
Summary
This summary is machine-generated.

RainbowSTORM is a new open-source software tool for analyzing spectroscopic single-molecule localization microscopy (sSMLM) data. This ImageJ/FIJI plugin enables quantitative spectral analysis and visualization for sSMLM imaging.

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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.