Attenuated Total Reflectance

Attenuated total reflectance (ATR) is a spectroscopic sampling technique that analyzes a material from its surface, often without extensive preparation, making it valuable for rapid chemical and materials characterization in engineering. In ATR Fourier-transform infrared spectroscopy, infrared light undergoes total internal reflection inside a high-refractive-index crystal; the resulting evanescent wave extends a short distance beyond the crystal surface and is attenuated when the sample absorbs specific infrared frequencies. The measured spectrum reveals molecular vibrations and chemical composition. ATR supports identification and comparison of polymers, coatings, composites, liquids, and contaminants, helping engineers assess material quality, surface treatments, degradation, and process performance.

Attenuated Total Reflectance - Related Videos

Education

JoVE Core - Analytical Chemistry

Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview

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2024

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...

Research

JoVE EoE - Neuroimaging

Cell Imaging Using Total Internal Reflection Fluorescence Microscopy

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2025

Source: Daniele, F., et. al TIRFM and pH-sensitive GFP-probes to Evaluate Neurotransmitter Vesicle Dynamics in SH-SY5Y Neuroblastoma Cells: Cell Imaging and Data Analysis. J. Vis. Exp. (2015).This video demonstrates the imaging of human neuroblastoma cells using a total internal reflection fluorescence (TIRF) microscope to visualize fluorophore-tagged synaptic vesicles. The cells are first focused in epifluorescence mode. Adjustments are then made to achieve total internal reflection,...

Total Internal Reflection Fluorescence Microscopy

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2023

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.

Total Internal Reflection Fluorescence Microscopy for Visualization of Exocytic Events

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2025

Source: Winkle, C. C., et al. Utilizing Combined Methodologies to Define the Role of Plasma Membrane Delivery During Axon Branching and Neuronal Morphogenesis. J. Vis. Exp. (2016)This video demonstrates total internal reflection fluorescence (TIRF) microscopy for visualizing exocytic events in cortical neurons. A pH-sensitive green fluorescent protein (GFP) marker fluoresces upon vesicle fusion, with selective TIRF excitation capturing membrane-proximal exocytosis in time-lapse imaging.

Detection and Quantification of Plasmodium falciparum in Aqueous Red Blood Cells by Attenuated Total Reflection Infrared Spectroscopy and Multivariate Data Analysis

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Cited by 1 •

2018

Here, we present a protocol for the detection and quantification of Plasmodium falciparum in infected aqueous red blood cells using an attenuated total reflection infrared spectrometer and multivariate data analysis.

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