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

Super-resolution Fluorescence Microscopy01:37

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.
Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
Fluorescence and Phosphorescence: Instrumentation01:25

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

Zeitschrift fur Rheumatologie·2021
Same author

Characterization of an Aux/IAA cDNA upregulated in Pinus pinaster roots in response to colonization by the ectomycorrhizal fungus Hebeloma cylindrosporum.

The New phytologist·2021
Same author

[Association of physical activity with fatigue and functional capacity in patients with rheumatoid arthritis].

Zeitschrift fur Rheumatologie·2020
Same author

[Severe polyneuropathy in primary Sjögren's syndrome : Sjögren's syndrome should be considered in patients with motor neuropathy].

Zeitschrift fur Rheumatologie·2020
Same author

Pollen-derived nonallergenic substances enhance Th2-induced IgE production in B cells.

Allergy·2015
Same author

Pollen-derived adenosine is a necessary cofactor for ragweed allergy.

Allergy·2015
Same journal

Multifunctional reconfigurable terahertz metasurface based on vanadium dioxide phase transition: achieving broadband absorption and efficient polarization conversion.

Applied optics·2026
Same journal

High-Q-factor electromagnetically induced transparency utilizing quasi-bound states in the continuum in an all-dielectric terahertz metasurface.

Applied optics·2026
Same journal

Automated stitching interferometry for high-precision metrology of X-ray mirrors.

Applied optics·2026
Same journal

Experimental demonstration of an approach to designing a metal-dielectric DBR resonant cavity structure.

Applied optics·2026
Same journal

High-precision wavefront reconstruction from a single-shot interferogram using a physics-driven hybrid feature calibration network.

Applied optics·2026
Same journal

Ultra-high-Q Fano resonance based on coupled topological corner states in Kagome photonic crystals.

Applied optics·2026
See all related articles

Related Experiment Video

Updated: Jun 16, 2026

Quantifying X-Ray Fluorescence Data Using MAPS
14:58

Quantifying X-Ray Fluorescence Data Using MAPS

Published on: February 17, 2018

Correction for light absorption within the object for quantitative microfluorometry: a theoretical approach.

D Ernst

    Applied Optics
    |January 30, 2010
    PubMed
    Summary
    This summary is machine-generated.

    Quantitative microfluorescence measurements can be complicated by light absorption. This study presents a simple method to test for and correct absorption errors in fluorescence intensity measurements.

    More Related Videos

    A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts
    08:43

    A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

    Published on: December 1, 2018

    Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy
    09:30

    Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

    Published on: January 18, 2017

    Related Experiment Videos

    Last Updated: Jun 16, 2026

    Quantifying X-Ray Fluorescence Data Using MAPS
    14:58

    Quantifying X-Ray Fluorescence Data Using MAPS

    Published on: February 17, 2018

    A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts
    08:43

    A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

    Published on: December 1, 2018

    Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy
    09:30

    Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

    Published on: January 18, 2017

    Area of Science:

    • * Optical microscopy
    • * Photometry
    • * Biophysical chemistry

    Background:

    • * Quantitative microfluorescence measurements typically integrate light intensity across the field of view.
    • * Light absorption by fluorescent dyes complicates intensity measurements, making them nonlinear functions of material concentration.
    • * The Lambert-Beer law describes the relationship between absorption and concentration, but its application in microfluorescence requires careful consideration.

    Purpose of the Study:

    • * To develop a straightforward method for assessing light absorption in quantitative microfluorescence.
    • * To provide formulas for correcting fluorescence intensity data affected by absorption.
    • * To enable accurate quantification of fluorescent materials despite absorption effects.

    Main Methods:

    • * Utilizing measurements at varying excitation wavelengths.
    • * Employing different excitation geometries.
    • * Analyzing data to detect deviations from linearity indicative of absorption.

    Main Results:

    • * A simple experimental approach to identify significant light absorption in microfluorescence.
    • * Formulas derived for correcting fluorescence intensity measurements affected by absorption.
    • * Demonstration that complex equipment is not required for absorption correction.

    Conclusions:

    • * Light absorption is a critical factor affecting quantitative microfluorescence accuracy.
    • * The described method offers a practical solution for testing and correcting absorption-induced errors.
    • * Accurate fluorescence quantification is achievable with simple adjustments to measurement protocols.