Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Fluorescence and Phosphorescence: Instrumentation

Fluorescence and Phosphorescence: Instrumentation

Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Evaluating Linker Architecture in RNA-Detecting Riboglow Probes and Effects on Fluorescence Turn-On.

ACS chemical biology·2026

Same author

Expanding the Riboglow-FLIM Toolbox with Different Fluorescence Lifetime-Producing RNA Tags.

Biochemistry·2025

Same author

Evaluating Riboglow-FLIM probes for RNA sensing.

RSC chemical biology·2024

Same author

Establishing Riboglow-FLIM to visualize noncoding RNAs inside live zebrafish embryos.

Biophysical reports·2023

Same author

It's complicated: the interplay of <i>Kif1c</i> mRNA localization in cell protrusions, assembly of protein binding partners on the KIF1C protein, and cell migration.

Genes & development·2023

Same author

Visualizing orthogonal RNAs simultaneously in live mammalian cells by fluorescence lifetime imaging microscopy (FLIM).

Nature communications·2023

Same journal

Mechanisms underpinning chromosome structure in metazoans.

Molecular biology of the cell·2026

Same journal

Conserved and Divergent Modes of Substrate Interaction Define Selective Localizations and Functions of a Cdc14 Phosphatase.

Molecular biology of the cell·2026

Same journal

Dimerization of the centriolin-like protein Nud1 governs spindle pole body inheritance in budding yeast.

Molecular biology of the cell·2026

Same journal

Non-muscle Myosin II acts as a negative feedback mediator to control cell contraction dynamics in adherent cells.

Molecular biology of the cell·2026

Same journal

The tetraspanin disc proteins, peripherin-2 and ROM1, facilitate CNG channel localization to the rod outer segment.

Molecular biology of the cell·2026

Same journal

Csf1 facilitates adaptive membrane lipid remodeling linked to ER-plasma membrane contact sites.

Molecular biology of the cell·2026

See all related articles

Search research articles

Related Experiment Video

Updated: May 4, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Practical guide to fluorescence lifetime imaging microscopy.

Nadia Sarfraz¹, Esther Braselmann¹

¹Department of Chemistry, Georgetown University, Washington, DC 20057.

Molecular Biology of the Cell

|May 27, 2025

Summary

This summary is machine-generated.

Fluorescence lifetime imaging microscopy (FLIM) offers advanced cellular analysis by sensing molecular environments and enabling RNA quantification. This guide empowers researchers to utilize FLIM for diverse biosensors and microscopy applications.

More Related Videos

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Published on: June 7, 2019

Fluorescence Lifetime Macro Imager for Biomedical Applications

Fluorescence Lifetime Macro Imager for Biomedical Applications

Published on: April 7, 2023

Related Experiment Videos

Last Updated: May 4, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Published on: June 7, 2019

Fluorescence Lifetime Macro Imager for Biomedical Applications

Fluorescence Lifetime Macro Imager for Biomedical Applications

Published on: April 7, 2023

Area of Science:

Biophotonics and advanced microscopy techniques.

Background:

Fluorescence lifetime imaging microscopy (FLIM) provides unique advantages over traditional intensity-based methods, including sensitivity to molecular environments and concentration independence.
FLIM has been recently adapted for quantitative RNA sensing in live cells, creating the Riboglow-FLIM platform.
The technology holds significant potential to complement and enhance existing microscopy platforms.

Purpose of the Study:

To provide a practical guide for utilizing FLIM with diverse fluorescence sensors.
To introduce researchers to the fundamental concepts and best practices for designing and executing FLIM experiments.
To demonstrate the broad applicability of FLIM workflows through real-world case studies.

Main Methods:

Discussion of FLIM basics, including principles of operation and data acquisition.
Explanation of experimental setup considerations for FLIM.
Overview of data-fitting principles and analysis techniques relevant to FLIM data.

Main Results:

Demonstration of FLIM's capability for quantitative RNA sensing in live cells using the Riboglow-FLIM platform.
Case studies showcasing the application of FLIM with well-characterized fluorescent proteins.
Validation of FLIM workflows for diverse fluorescence sensing applications.

Conclusions:

FLIM is a powerful modality for sensing molecular environments, binding events, and metabolism.
The Riboglow-FLIM platform represents a significant advancement for quantitative RNA analysis in live cells.
This guide equips researchers with the knowledge to effectively implement FLIM for various biological sensing applications.