Abstract
SUMMARY: We introduce software for reading, writing and processing fluorescence single-molecule and image spectroscopy data and developing analysis pipelines to unify various spectroscopic analysis tools. Our software can be used for processing multiple experiment types, e.g. for time-resolved single-molecule spectroscopy, laser scanning microscopy, fluorescence correlation spectroscopy and image correlation spectroscopy. The software is file format agnostic and processes multiple time-resolved data formats and outputs. Our software eliminates the need for data conversion and mitigates data archiving issues. AVAILABILITY AND IMPLEMENTATION: tttrlib is available via pip (https://pypi.org/project/tttrlib/) and bioconda while the open-source code is available via GitHub (https://github.com/fluorescence-tools/tttrlib). Presented examples and additional documentation demonstrating how to implement in vitro and live-cell image spectroscopy analysis are available at https://docs.peulen.xyz/tttrlib and https://zenodo.org/records/14002224.