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

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.
Interference and Diffraction02:18

Interference and Diffraction

Interference is a characteristic phenomenon exhibited by waves. When two electromagnetic waves interact with their peaks and troughs coinciding, a resulting wave with enhanced amplitude is produced. This is known as constructive interference. In this case, the two waves interacting are in phase with each other.
Phase Contrast and Differential Interference Contrast Microscopy01:26

Phase Contrast and Differential Interference Contrast Microscopy

Phase-Contrast Microscopes
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to create high-contrast, high-resolution images without staining. It is the oldest and simplest type of microscope that creates an image by altering the wavelengths of light rays passing through the specimen. Altered wavelength paths are created using an annular stop in the condenser. The annular stop produces a hollow cone of...
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.
Three-Dimensional Microscopy in Microbiology01:28

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...
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...

You might also read

Related Articles

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

Sort by
Same author

Neutrophil degranulation and extracellular ROS production are inactivated by Yersinia pseudotuberculosis YopE through a SKAP2 independent pathway.

bioRxiv : the preprint server for biology·2026
Same author

Differentiating <i>Peromyscus leucopus</i> bone marrow-derived macrophages for characterization of responses to <i>Borrelia burgdorferi</i> and lipopolysaccharide.

Infection and immunity·2025
Same author

STING promotes homeostatic maintenance of tissues and confers longevity with aging.

bioRxiv : the preprint server for biology·2024
Same author

Heterogeneity of Signaling Complex Nanostructure in T Cells Activated Via the T Cell Antigen Receptor.

Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·2023
Same author

Inherited ARPC5 mutations cause an actinopathy impairing cell motility and disrupting cytokine signaling.

Nature communications·2023
Same author

Neutrophils require SKAP2 for reactive oxygen species production following C-type lectin and <i>Candida</i> stimulation.

iScience·2021
Same journal

Measuring Mitochondrial Respiration in Previously Frozen Biological Samples.

Current protocols in cell biology·2020
Same journal

Proximity Ligation Assay for Detecting Protein-Protein Interactions and Protein Modifications in Cells and Tissues in Situ.

Current protocols in cell biology·2020
Same journal

Methods for Investigating Corneal Cell Interactions and Extracellular Vesicles In Vitro.

Current protocols in cell biology·2020
Same journal

Multiplexed Proximity Biotinylation Coupled to Mass Spectrometry for Defining Integrin Adhesion Complexes.

Current protocols in cell biology·2020
Same journal

Preparation of Extracellular Matrix Paper and Construction of Multi-Layered 3D Tissue Model.

Current protocols in cell biology·2020
Same journal

Metabolic Analysis at the Nanoscale with Multi-Isotope Imaging Mass Spectrometry (MIMS).

Current protocols in cell biology·2020
See all related articles

Related Experiment Video

Updated: Jun 17, 2026

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules
09:45

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules

Published on: August 8, 2019

Interference reflection microscopy.

Valarie A Barr1, Stephen C Bunnell

  • 1National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.

Current Protocols in Cell Biology
|December 17, 2009
PubMed
Summary
This summary is machine-generated.

Interference reflection microscopy (IRM) provides high-contrast cell imaging on glass surfaces. This technique is valuable for studying cell adhesion and mobility, especially when combined with laser scanning confocal microscopy.

More Related Videos

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins
06:43

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Published on: May 3, 2022

Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy
10:19

Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy

Published on: January 21, 2019

Related Experiment Videos

Last Updated: Jun 17, 2026

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules
09:45

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules

Published on: August 8, 2019

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins
06:43

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Published on: May 3, 2022

Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy
10:19

Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy

Published on: January 21, 2019

Area of Science:

  • Biophysics
  • Cell Biology
  • Optical Microscopy

Background:

  • Interference reflection microscopy (IRM) is an optical technique.
  • It generates high-contrast images by utilizing the interference of reflected light waves.
  • IRM is particularly useful for studying cell-substratum interactions.

Purpose of the Study:

  • To present methods for obtaining interference reflection microscopy (IRM) images of cells.
  • To emphasize IRM imaging using a laser scanning confocal microscope (LSCM).
  • To detail techniques for imaging both fixed and live cells.

Main Methods:

  • Utilizing the interference of reflected light waves to create high-contrast images.
  • Adapting laser scanning confocal microscopes (LSCM) for IRM imaging without instrument modification.
  • Implementing techniques for simultaneous multi-channel capture of fluorescence and reflection images.

Main Results:

  • High-contrast and high-definition images of cells on glass coverslips.
  • Successful application of IRM for studying cell adhesion and mobility.
  • Demonstration of IRM compatibility with LSCM for enhanced imaging capabilities.

Conclusions:

  • IRM is a versatile technique for visualizing cell-substratum interactions.
  • LSCM can be readily used for IRM imaging, often without modifications.
  • The methods presented allow for comprehensive imaging of cellular dynamics and structures.