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.
Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview01:13

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...

You might also read

Related Articles

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

Sort by
Same author

Retention rate of a novel autoinjector e-Device introduced to patients with chronic arthritis treated with certolizumab pegol in clinical practice: an observational implementation study.

Scandinavian journal of rheumatology·2024
Same author

Decompressive craniectomy to cranioplasty: a retrospective observational study using Hospital Episode Statistics in England.

BMJ surgery, interventions, & health technologies·2024
Same author

Increased prevalence of sleep disturbance in psoriatic arthritis is associated with inflammatory and non-inflammatory measures.

Scandinavian journal of rheumatology·2022
Same author

The reliability of pseudoneglect is task dependent.

Neuropsychologia·2020
Same author

Pattern contrast influences wariness in naïve predators towards aposematic patterns.

Scientific reports·2020
Same author

Whole-genome sequencing reveals clinically relevant insights into the aetiology of familial breast cancers.

Annals of oncology : official journal of the European Society for Medical Oncology·2019
Same journal

Heterojunction Gate-Empowered OPECT Aptasensing: A Valid Protocol for Realizing High Current Gain at Low Electron Donor Dependency.

Analytical chemistry·2026
Same journal

Development of a Tapping-Mode Scanning Probe Electrospray Ionization Platform for High-Sensitivity and Long-Term Stability in Single-Cell Mass Spectrometry Imaging of Tissue.

Analytical chemistry·2026
Same journal

A Solid-State Near-Infrared Fluorescent Probe by a Synergistic Extended Conjugated System for Detecting Cys with Long-Term Imaging in Orthotopic Bladder Cancer.

Analytical chemistry·2026
Same journal

Air-Based Porous Array Dielectric Barrier Discharge Ionization Source for Explosive Trace Detection.

Analytical chemistry·2026
Same journal

PSAQ<sup>+1</sup>: Absolute Protein Quantification Using a <sup>13</sup>C<sub>1</sub>-Labeled Protein Standard, Coisolation of Peptide Pairs and LC-PRM.

Analytical chemistry·2026
Same journal

Heterojunction-Enhanced Interfacial Evanescent-Tunable Fiber Optic Probe for Amplification-free CRISPR/Cas12a-Based Rapid and Ultrasensitive Detection of MPXV.

Analytical chemistry·2026
See all related articles

Related Experiment Video

Updated: Jun 1, 2026

Total Internal Reflection Absorption Spectroscopy (TIRAS) for the Detection of Solvated Electrons at a Plasma-liquid Interface
08:50

Total Internal Reflection Absorption Spectroscopy (TIRAS) for the Detection of Solvated Electrons at a Plasma-liquid Interface

Published on: January 24, 2018

Total internal reflection fluorescence correlation spectroscopy for counting molecules at solid/liquid interfaces.

R L Hansen1, J M Harris

  • 1Department of Chemistry, University of Utah, Salt Lake City, Utah 84112.

Analytical Chemistry
|June 8, 2011
PubMed
Summary
This summary is machine-generated.

This study introduces total internal reflection fluorescence correlation spectroscopy (TIR-FCS) for quantifying molecules at interfaces. This method accurately determines molecular populations without calibration, even with low signal detection.

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

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)
09:14

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)

Published on: October 2, 2012

Related Experiment Videos

Last Updated: Jun 1, 2026

Total Internal Reflection Absorption Spectroscopy (TIRAS) for the Detection of Solvated Electrons at a Plasma-liquid Interface
08:50

Total Internal Reflection Absorption Spectroscopy (TIRAS) for the Detection of Solvated Electrons at a Plasma-liquid Interface

Published on: January 24, 2018

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

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)
09:14

Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)

Published on: October 2, 2012

Area of Science:

  • Analytical Chemistry
  • Surface Science
  • Spectroscopy

Background:

  • Accurate quantification of molecular populations at solid/liquid interfaces is crucial for understanding surface phenomena.
  • Existing methods often require sensitivity calibration, standards, or complex kinetic modeling.
  • Single-molecule counting can be limited by low signal detection per molecule.

Purpose of the Study:

  • To develop and validate total internal reflection fluorescence correlation spectroscopy (TIR-FCS) as a quantitative method for interfacial molecular populations.
  • To demonstrate TIR-FCS's ability to quantify molecules without calibration or kinetic fitting.
  • To investigate the adsorption of rhodamine 6G dye molecules onto C-18-modified silica surfaces.

Main Methods:

  • Utilized total internal reflection fluorescence correlation spectroscopy (TIR-FCS) to monitor fluorescence signal fluctuations at solid/liquid interfaces.
  • Applied Poisson statistics to analyze the low-frequency noise in the fluorescence signal, correlating it to molecular origins.
  • Measured surface populations of rhodamine 6G dye on C-18-derivatized silica and compared results with chromatographic data.

Main Results:

  • TIR-FCS successfully quantified molecular populations at the interface by analyzing noise magnitude via Poisson statistics.
  • Quantitative results were obtained without the need for sensitivity calibration or kinetic model fitting.
  • The method provided accurate measurements even with low photoelectron detection per molecule.

Conclusions:

  • TIR-FCS is a robust and quantitative technique for determining molecular numbers at solid/liquid interfaces.
  • The method offers advantages over traditional techniques by simplifying experimental requirements and enhancing applicability.
  • Adsorption characteristics, including free energy contributions, of rhodamine 6G on modified silica surfaces were examined.