Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Silicon Plasmonics for Enhanced Responsivity of Silicon Photodetectors in Deep-Ultraviolet Region.

Physical review letters·2025
Same author

Non-invasive nanoscale imaging of protein micro- and nanocrystals for screening crystallization conditions.

Journal of applied crystallography·2024
Same author

Development of a localized surface plasmon-enhanced electron beam-pumped nanoscale light source for electron beam excitation-assisted optical microscopy.

Microscopy (Oxford, England)·2024
Same author

High resolution imaging of ultrafine bubbles in water by Atmospheric SEM-CL.

Micron (Oxford, England : 1993)·2022
Same author

A Case of Intraocular Lymphoma Diagnosed by Subretinal Fluid Biopsy.

International medical case reports journal·2022
Same author

High Spatial Resolution Ion Imaging by Focused Electron-Beam Excitation with Nanometric Thin Sensor Substrate.

Sensors (Basel, Switzerland)·2022

Related Experiment Video

Updated: Dec 29, 2025

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy
10:41

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

Published on: June 7, 2019

8.9K

Ultrashort laser based two-photon phase-resolved fluorescence lifetime measurement method.

Hafizah Halip1, Yuto Yoshimura2, Wataru Inami1,3

  • 1Graduate School of Science and Technology, Shizuoka University, 3-5-1 Johoku, Naka, Hamamatsu 432-8561, Japan.

Methods and Applications in Fluorescence
|January 31, 2020
PubMed
Summary

This study introduces a new two-photon fluorescence-lifetime measurement technique using a simple laser setup. The method accurately determines fluorescence lifetimes, validated with rhodamine B and a cellular thermoprobe.

More Related Videos

High Resolution Phonon-assisted Quasi-resonance Fluorescence Spectroscopy
10:40

High Resolution Phonon-assisted Quasi-resonance Fluorescence Spectroscopy

Published on: June 28, 2016

7.9K
An Experimental Protocol for Femtosecond NIR/UV - XUV Pump-Probe Experiments with Free-Electron Lasers
09:49

An Experimental Protocol for Femtosecond NIR/UV - XUV Pump-Probe Experiments with Free-Electron Lasers

Published on: October 23, 2018

16.4K

Related Experiment Videos

Last Updated: Dec 29, 2025

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy
10:41

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

Published on: June 7, 2019

8.9K
High Resolution Phonon-assisted Quasi-resonance Fluorescence Spectroscopy
10:40

High Resolution Phonon-assisted Quasi-resonance Fluorescence Spectroscopy

Published on: June 28, 2016

7.9K
An Experimental Protocol for Femtosecond NIR/UV - XUV Pump-Probe Experiments with Free-Electron Lasers
09:49

An Experimental Protocol for Femtosecond NIR/UV - XUV Pump-Probe Experiments with Free-Electron Lasers

Published on: October 23, 2018

16.4K

Area of Science:

  • Optics and Photonics
  • Biophysical Chemistry

Background:

  • Fluorescence lifetime measurements are crucial for understanding molecular dynamics and environmental sensing.
  • Existing methods can be complex or require specialized equipment.

Purpose of the Study:

  • To develop and validate a simplified two-photon phase-resolved fluorescence-lifetime measurement method.
  • To demonstrate the method's applicability for biological and chemical sensing.

Main Methods:

  • Utilized a two-photon excitation scheme with an ultrashort pulse laser.
  • Employed a lock-in amplifier to precisely control phase differences between reference and fluorescence signals.
  • Assumed monoexponential fluorescence decay for all measurements.

Main Results:

  • Accurately measured the fluorescence lifetime of rhodamine B as 2.15 ns.
  • Quantified the temperature sensitivity of a cellular thermoprobe at 1.39 ns/°C within a physiological temperature range (33.79–37.2 °C).

Conclusions:

  • The proposed two-photon phase-resolved method offers a feasible and accurate approach for fluorescence lifetime determination.
  • The technique's simplicity and accuracy make it suitable for various applications, including temperature sensing in biological systems.