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

Total Internal Reflection Fluorescence Microscopy

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.

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.

You might also read

Related Articles

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

Sort by

Same author

Quantitative comparison of methodologies for translation site imaging in living cells.

RNA (New York, N.Y.)·2026

Same author

An integrated RNA-centric imaging and omics approach reveals distinct properties and composition of neuronal RNA granules.

bioRxiv : the preprint server for biology·2026

Same author

RNA-binding proteins in the mouse lens: Functional classifications, expression profiling, and interaction studies of Carhsp1 with crystallin mRNAs.

Developmental biology·2026

Same author

Single-mRNA imaging and modeling reveal coupled translation initiation and elongation rates.

eLife·2026

Same author

A series of spontaneously blinking dyes for super-resolution microscopy.

Nature methods·2026

Same author

A platform for analyzing translational control by RBPs at single-mRNA resolution in cells.

Biophysics and physicobiology·2026

Same journal

Evolutionary and Biochemical Perspectives on the Incorporation and Utilization of Selenocysteine.

Cold Spring Harbor perspectives in biology·2026

Same journal

The Mitochondrial Calcium Uniporter: From Parts to Signaling Networks.

Cold Spring Harbor perspectives in biology·2026

Same journal

Growth Control and Beyond: Functional Diversity and Regulation of the Hippo Pathway in the Nervous System.

Cold Spring Harbor perspectives in biology·2026

Same journal

Structural Studies of Core Hippo Pathway Components.

Cold Spring Harbor perspectives in biology·2026

Same journal

The Hippo Pathway in Intestinal Regeneration, Fetal Reprogramming, and Tumorigenesis.

Cold Spring Harbor perspectives in biology·2026

Same journal

A Synergy between Genetics and Biochemistry Unravels the Molecular Architecture of the Hippo Signaling Pathway.

Cold Spring Harbor perspectives in biology·2026

See all related articles

Search research articles

Related Experiment Video

Updated: May 18, 2026

An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics

An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics

Published on: September 15, 2020

Imaging translation in single cells using fluorescent microscopy.

Jeffrey A Chao¹, Young J Yoon, Robert H Singer

¹Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.

Cold Spring Harbor Perspectives in Biology

|September 11, 2012

Summary

This summary is machine-generated.

Researchers can now visualize protein synthesis in single cells using advanced imaging techniques. This allows for a deeper understanding of how and where proteins are made, offering new insights into cellular regulation.

More Related Videos

Quantitative Immunofluorescence to Measure Global Localized Translation

Quantitative Immunofluorescence to Measure Global Localized Translation

Published on: August 22, 2017

Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)

Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)

Published on: March 15, 2013

Related Experiment Videos

Last Updated: May 18, 2026

An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics

An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics

Published on: September 15, 2020

Quantitative Immunofluorescence to Measure Global Localized Translation

Quantitative Immunofluorescence to Measure Global Localized Translation

Published on: August 22, 2017

Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)

Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)

Published on: March 15, 2013

Area of Science:

Molecular Biology
Cell Biology
Neuroscience

Background:

Translational control regulates protein abundance, timing, and location.
Traditional methods measure average protein synthesis, obscuring cellular details.
Fluorescent microscopy enables studying translation within the cellular context.

Purpose of the Study:

Highlight recent methodologies for studying translation in single cells.
Enable investigation of global protein synthesis changes.
Facilitate the study of specific mRNA regulation.

Main Methods:

Fluorescent microscopy techniques and reagents.
Single-cell analysis of protein synthesis.
Imaging of messenger RNA (mRNA) translation.

Main Results:

Recent methodologies allow for studying global protein synthesis in single cells.
Specific mRNA regulation can be investigated at the single-cell level.
Direct evidence for local mRNA translation at neuronal synapses was observed.

Conclusions:

Single-cell imaging provides mechanistic insights into translational control.
This approach overcomes limitations of ensemble measurements.
Imaging translation is a powerful tool for studying cellular regulation and neuroscience.