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.

Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview

Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview

Attenuated total reflectance (ATR) infrared spectroscopy is a powerful analytical technique used to study the composition of materials. It is widely employed in chemistry, materials science, forensic science, and other fields where sample characterization is required. ATR has several advantages over traditional transmission IR spectroscopy, including the requirement of little to no sample preparation and the ability to analyze a wide range of samples.
The ATR process begins by directing a beam...

Fluorescence and Phosphorescence: Instrumentation

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

Controlling Microarray Feature Spreading and Response Stability on Porous Silicon Platforms by Using Alkene-Terminal Ionic Liquids and UV Hydrosilylation.

Langmuir : the ACS journal of surfaces and colloids·2020

Same author

Interplay Between Silicon Nanocrystal Size and Local Environment Within Porous Silicon on the Analyte-Dependent Photoluminescence Response.

Applied spectroscopy·2019

Same author

Effects of Acetone Vapor on the Exciton Band Photoluminescence Emission from Single- and Few-Layer WS<sub>2</sub> on Template-Stripped Gold.

Sensors (Basel, Switzerland)·2019

Same author

Filterless optical oxygen sensor based on a CMOS buried double junction photodiode.

Sensors and actuators. B, Chemical·2018

Same author

Gallium indium eutectic masking prior to porous silicon formation creates unique spatially-dependent chemistries.

Analytica chimica acta·2018

Same author

Exploiting the 3-Aminopropyltriethoxysilane (APTES) autocatalytic nature to create bioconjugated microarrays on hydrogen-passivated porous silicon.

Talanta·2017

Search research articles

Related Experiment Video

Updated: May 27, 2026

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

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

Published on: May 3, 2022

High-pressure total internal reflection fluorescence apparatus.

Michael J Dabney¹, Frank V Bright

¹Department of Chemistry, Natural Sciences Complex, University at Buffalo, The State University of New York, Buffalo, New York 14260-3000, USA.

Applied Spectroscopy

|November 8, 2011

Summary

This summary is machine-generated.

A new high-pressure total internal reflection fluorescence (HP-TIRF) apparatus enables rapid measurements of interfacial dynamics. Fluorinated silane monolayers show altered microviscosity in supercritical carbon dioxide due to solubility effects.

More Related Videos

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

Published on: March 22, 2012

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

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: May 27, 2026

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

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

Published on: May 3, 2022

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins

Published on: March 22, 2012

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

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

Published on: January 21, 2019

Area of Science:

Surface science
Physical chemistry
Materials science

Background:

Interfacial processes are critical in diverse fields like medicine and separations science.
These processes occur under various conditions, necessitating advanced measurement techniques.

Purpose of the Study:

To introduce a novel high-pressure total internal reflection fluorescence (HP-TIRF) apparatus.
To investigate the influence of supercritical carbon dioxide on interfacial dynamics.

Main Methods:

Utilized a new HP-TIRF system for rapid fluorescence measurements up to 250 bar and 293-353 K.
Studied rotational reorientation dynamics of BODIPY 494/503 on silanized silica surfaces in supercritical carbon dioxide.
Employed the Debye-Stokes-Einstein expression to analyze dynamics.

Main Results:

Observed a decrease in local microviscosity around BODIPY 494/503 as supercritical carbon dioxide density increased.
Found that the terminal group of the silane monolayer (methyl vs. trifluoromethyl) significantly impacts viscosity changes.
Demonstrated that fluorine-containing species' solubility in supercritical carbon dioxide explains the observed effects.

Conclusions:

The HP-TIRF apparatus provides picosecond time resolution for interfacial studies under high pressure.
Silane monolayer terminal groups and supercritical carbon dioxide density are key factors influencing interfacial microviscosity.
Solubility of fluorinated compounds in supercritical carbon dioxide plays a crucial role in interfacial behavior.