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Infrared (IR) Spectroscopy: Overview

Infrared (IR) Spectroscopy: Overview

When electromagnetic radiation passes through a material, atoms or molecules transition from a lower to a higher energy state by absorbing radiation corresponding to the energy difference between the two states. The absorption of infrared (IR) radiation causes transitions between vibrational energy levels in a molecule. Therefore, IR spectroscopy is a useful analytical tool for determining the molecular structure of molecules.
Different compounds display unique properties due to their...

IR Spectrum

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When infrared (IR) radiation passes through a molecule, the bonds stretch or bend by absorbing the radiation. This absorption creates the molecule's absorption spectrum, which is the plot of its percentage transmittance versus wavenumber.
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There are two main infrared (IR) spectrophotometers: dispersive IR spectrometers and Fourier transform infrared (FTIR) spectrometers. In a dispersive IR spectrometer, a beam of infrared radiation produced by a hot wire is divided into two parallel equal-intensity beams using mirrors. One beam passes through the sample, while another is a reference beam. The beams then move through the monochromator, which separates the radiations into a continuous spectrum of different frequencies. The...

IR Frequency Region: Fingerprint Region

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IR spectra are divided into two main regions: the diagnostic region and the fingerprint region. The diagnostic region of the spectrum lies above 1500 cm−1. The absorptions resulting from single-bond vibrations of the N–H, C–H, and O–H stretch at higher wavenumbers and appear on the left side of the spectrum. The stretching absorptions of the C≡C and C≡N occur between 2100–2300 cm−1. In contrast, those arising from stretching absorptions of the C=O, C=N, and C=C occur between 1600–1850 cm−1.
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The non-destructive nature and ability to provide valuable chemical information make IR spectroscopy a versatile technique with broad applications in various scientific and industrial fields. IR spectroscopy is commonly used to identify and characterize organic and inorganic compounds. It provides information about the functional groups present in a molecule and the bonding between atoms. This helps in the structural elucidation of compounds during organic synthesis, pharmaceutical research,...

Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview

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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.
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High-definition Fourier Transform Infrared (FT-IR) Spectroscopic Imaging of Human Tissue Sections towards Improving Pathology

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Published on: January 21, 2015

GLASSR-Net: Glass Substrate Spectral Restoration Neural Network for Fourier Transform Infrared Microspectroscopy in

Xiangyu Zhao¹, Jingzhu Shao¹, Yudong Tian¹

¹Center for Biophotonics, Institute of Medical Robotics, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China.

Analytical Chemistry

|February 26, 2025

Summary

Fourier transform infrared (FTIR) microspectroscopy can now be used with cost-effective glass slides. A new neural network, GLASSR-Net, restores spectral data, enabling wider clinical use of this pathology tool.

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Area of Science:

Biomedical Engineering
Spectroscopy
Computational Pathology

Background:

Fourier transform infrared (FTIR) microspectroscopy is a valuable tool for disease biomarker identification.
Current limitations include reliance on expensive and fragile IR-transparent substrates (e.g., CaF2/BaF2) or opaque IR-reflective substrates.
Pathological glass slides are cost-effective and standard in clinical settings, but not ideal for FTIR microspectroscopy.

Purpose of the Study:

To develop and validate a method for collecting high-quality FTIR spectra directly from standard glass pathological substrates.
To establish a computational approach for restoring spectral information lost when using glass slides.
To enable the integration of FTIR microspectroscopy into routine clinical histology workflows.

Main Methods:

Development of the glass substrate spectral restoration neural network (GLASSR-Net).
Acquisition of FTIR raster scanning data from contiguous tissue sections of papillary thyroid carcinoma (PTC) on both glass and CaF2 substrates.
Training and validation of GLASSR-Net using glass-based spectra as input and CaF2-based spectra as ground truth.

Main Results:

GLASSR-Net successfully restored fingerprint absorbance spectra (1800-1000 cm-1) from glass-based FTIR data.
The restored spectra accurately reconstructed biochemical distribution in both spatial and spectral domains.
Analysis of restored spectra revealed PTC-specific biochemical signatures, including decreased amide I/II absorption and increased lipids/nucleic acids in cancerous regions.

Conclusions:

GLASSR-Net provides a novel framework for spectral restoration in glass-based FTIR microspectroscopy.
This methodology overcomes substrate limitations, facilitating the clinical translation of FTIR microspectroscopy.
The approach integrates traditional spectral histology with advanced computational methods for enhanced pathological analysis.