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

Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.

Confocal Fluorescence Microscopy

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

Three-Dimensional Microscopy in Microbiology

Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...

You might also read

Related Articles

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

Sort by

Same author

Dysbiosis of skin microbiota with increased fungal diversity is associated with severity of disease in atopic dermatitis.

Journal of the European Academy of Dermatology and Venereology : JEADV·2022

Same author

In vitro efficacy of antifungal agents alone and in shampoo formulation against dandruff-associated Malassezia spp. and Staphylococcus spp.

International journal of cosmetic science·2019

Same author

[Cutaneous manifestations of Lyme disease : Pitfalls in the serological diagnostic workup].

Der Hautarzt; Zeitschrift fur Dermatologie, Venerologie, und verwandte Gebiete·2017

Same author

EAACI Molecular Allergology User's Guide.

Pediatric allergy and immunology : official publication of the European Society of Pediatric Allergy and Immunology·2016

Same author

Global Allergy Forum and 3rd Davos Declaration 2015: Atopic dermatitis/Eczema: challenges and opportunities toward precision medicine.

Allergy·2016

Same author

Tinea capitis and tinea faciei in the Zurich area - an 8-year survey of trends in the epidemiology and treatment patterns.

Journal of the European Academy of Dermatology and Venereology : JEADV·2014

Search research articles

Related Experiment Video

Updated: Jul 6, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Four-dimensional multiphoton microscopy with time-correlated single-photon counting.

A Schönle¹, M Glatz, S W Hell

¹High Resolution Optical Microscopy Group, Max-Planck-Institute for Biophysical Chemistry, D-37070 Göttingen, Germany.

|March 21, 2008

Summary

We developed a new fluorescence-lifetime imaging method using multiphoton microscopy and time-correlated single-photon counting. This technique precisely measures fluorescence lifetimes in biological samples and materials.

More Related Videos

Time-resolved Photophysical Characterization of Triplet-harvesting Organic Compounds at an Oxygen-free Environment Using an iCCD Camera

Time-resolved Photophysical Characterization of Triplet-harvesting Organic Compounds at an Oxygen-free Environment Using an iCCD Camera

Published on: December 27, 2018

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Related Experiment Videos

Last Updated: Jul 6, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Time-resolved Photophysical Characterization of Triplet-harvesting Organic Compounds at an Oxygen-free Environment Using an iCCD Camera

Time-resolved Photophysical Characterization of Triplet-harvesting Organic Compounds at an Oxygen-free Environment Using an iCCD Camera

Published on: December 27, 2018

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Area of Science:

Biophysics
Microscopy
Photonics

Background:

Multiphoton excitation microscopy offers deep tissue penetration and reduced phototoxicity.
Fluorescence lifetime imaging (FLIM) provides functional and molecular information beyond intensity.
Accurate and efficient FLIM implementation is crucial for advanced biological imaging.

Purpose of the Study:

To implement and validate a PC-compatible fluorescence-lifetime imaging system for multiphoton microscopy.
To enable high-resolution, four-dimensional data acquisition with detailed fluorescence decay analysis.
To demonstrate the system's capability in analyzing both material and biological samples.

Main Methods:

Utilized PC-compatible modules for time-correlated single-photon counting (TCSPC).
Acquired four-dimensional data stacks, with each pixel containing fluorescence decay curves (up to 4096 bins).
Employed statistical methods for extracting fluorescence lifetimes and amplitudes at pixel or region-of-interest levels.

Main Results:

Achieved a temporal response function width of 420 ps using an avalanche photodiode.
Demonstrated lifetime precision in the tens of picoseconds, dependent on signal-to-noise ratio.
Successfully visualized lifetime changes in fluorescent layers and Förster resonance energy transfer (FRET) in live mammalian cells.

Conclusions:

The implemented FLIM system provides precise fluorescence lifetime measurements.
The system is versatile, applicable to both material science and live-cell imaging.
This technology enhances multiphoton microscopy capabilities for advanced biological and material studies.