Embracing the power of fluorescence lifetime imaging

Jörg Enderlein

doi:10.1515/mim-2025-0003

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Embracing the power of fluorescence lifetime imaging

Jörg Enderlein

Published/Copyright: March 20, 2025

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From the journal Methods in Microscopy Volume 2 Issue 1

Welcome to this special edition of Methods in Microscopy (MiM), dedicated to the fascinating world of Fluorescence Lifetime Imaging (FLIM). As technology advances, FLIM has emerged as a powerful technique, offering unique insights into biological processes and material properties. In this issue, we explore the principles, applications, and recent advancements in FLIM, highlighting its growing impact on microscopy.

FLIM provides several advantages over traditional fluorescence intensity-based imaging. By measuring the decay rate of an excited state, it reveals crucial information about the local environment surrounding a fluorescent molecule. This enables researchers to study a wide range of phenomena, including protein interactions, ion concentrations, and Förster Resonance Energy Transfer (FRET).

One of FLIM’s key strengths is its ability to deliver quantitative data that is independent of fluorophore concentration and photobleaching effects. This makes it particularly useful for long-term imaging experiments and for comparing samples under varying conditions. Additionally, FLIM can be integrated with other imaging modalities, such as confocal microscopy and two-photon excitation, further enhancing its capabilities.

In this special edition, our guest editors have curated a collection of tutorials and reviews designed to serve as a comprehensive guide for both novice and experienced researchers. From the fundamental principles of FLIM to advanced techniques such as spectral FLIM and high-throughput FLIM, this issue covers a broad spectrum of topics to equip you with the knowledge and skills needed to harness the full potential of FLIM in your research.

We also provide practical guidance on data acquisition and analysis, along with real-world examples of successful FLIM applications across diverse fields, including cell biology, neuroscience, and materials science. By showcasing FLIM’s versatility, the editorial team of Methods in Microscopy hopes to inspire you to integrate this powerful technique into your own research endeavors.

Like any microscopy method, FLIM comes with its own set of challenges and limitations. However, continuous technological advancements and methodological developments are expanding its possibilities. With the advent of new fluorophores, improved detectors, and sophisticated data analysis tools, FLIM is becoming increasingly accessible and powerful.

I wish this journal success in bridging the gap between innovation and practical application. To support this mission, I encourage you to contribute your methods and share best practices with the community.

As a fellow researcher once stated: “The future of microscopy depends not only on technological advancements but also on ensuring that researchers can effectively apply them.”

Corresponding author: Jörg Enderlein, Third Institute of Physics – Biophysics, Georg-August-Universitat Gottingen, Friedrich-Hund-Platz 1, 37073 Göttingen, Germany, E-mail: jenderl@gwdg.de

Published Online: 2025-03-20

Published in Print: 2025-04-28

This work is licensed under the Creative Commons Attribution 4.0 International License.

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https://doi.org/10.1515/mim-2025-0003

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