Identification of a Glass Substrate to Study Cells Using Fourier Transform Infrared Spectroscopy: Are We Closer to Spectral Pathology?

The rising incidence of cancer worldwide is causing an increase in the workload in pathology departments. This, coupled with advanced analysis methodologies, supports a developing need for techniques that could identify the presence of cancer cells in cytology and tissue samples in an objective, fast, and automated way. Fourier transform infrared (FT-IR) microspectroscopy can identify cancer cells in such samples objectively. Thus, it has the potential to become another tool to help pathologists in their daily work. However, one of the main drawbacks is the use of glass substrates by pathologists. Glass absorbs IR radiation, removing important mid-IR spectral data in the fingerprint region (1800 cm−1 to 900 cm−1). In this work, we hypothesized that, using glass coverslips of differing compositions, some regions within the fingerprint area could still be analyzed. We studied three different types of cells (peripheral blood mononuclear cells, a leukemia cell line, and a lung cancer cell line) and lymph node tissue placed on four different types of glass coverslips. The data presented here show that depending of the type of glass substrate used, information within the fingerprint region down to 1350 cm−1 can be obtained. Furthermore, using principal component analysis, separation between the different cell lines was possible using both the lipid region and the fingerprint region between 1800 cm−1 and 1350 cm−1. This work represents a further step towards the application of FT-IR microspectroscopy in histopathology departments.

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In Vitro Spectroscopy-Based Profiling of Urothelial Carcinoma: A Fourier Transform Infrared and Raman Imaging Study

Cancers ◽

10.3390/cancers13010123 ◽

2021 ◽

Vol 13 (1) ◽

pp. 123

Author(s):

Monika Kujdowicz ◽

Wojciech Placha ◽

Brygida Mech ◽

Karolina Chrabaszcz ◽

Krzysztof Okoń ◽

...

Keyword(s):

Bladder Cancer ◽

Fourier Transform ◽

Cancer Cells ◽

Cell Line ◽

Cell Lines ◽

Malignant Neoplasm ◽

Fourier Transform Infrared ◽

Diagnostic Tools ◽

Label Free ◽

Bladder Cancer Cells

Markers of bladder cancer cells remain elusive, which is a major cause of the low recognition of this malignant neoplasm and its recurrence. This implies an urgent need for additional diagnostic tools which are based on the identification of the chemism of bladder cancer. In this study, we employed label-free techniques of molecular imaging—Fourier Transform Infrared and Raman spectroscopic imaging—to investigate bladder cancer cell lines of various invasiveness (T24a, T24p, HT-1376, and J82). The urothelial HCV-29 cell line was the healthy control. Specific biomolecules discriminated spatial distribution of the nucleus and cytoplasm and indicated the presence of lipid bodies and graininess in some cell lines. The most prominent discriminators are the total content of lipids and sugar moieties as well as the presence of glycogen and other carbohydrates, un/saturated lipids, cytochromes, and a level of S-S bridges in proteins. The combination of the obtained hyperspectral database and chemometric methods showed a clear differentiation of each cell line at the level of the nuclei and cytoplasm and pointed out spectral signals which differentiated bladder cancer cells. Registered spectral markers correlated with biochemical composition changes can be associated with pathogenesis and potentially used for the diagnosis of bladder cancer and response to experimental therapies.

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Fourier Transform Infrared (FT-IR) Microspectroscopy of 20th Century Russian Oil Paintings: Problem of Dating

Applied Spectroscopy ◽

10.1177/0003702816652330 ◽

2016 ◽

Vol 70 (7) ◽

pp. 1150-1156 ◽

Cited By ~ 6

Author(s):

Irina A. Balakhnina ◽

Nikolay N. Brandt ◽

Andrey Yu Chikishev ◽

Yurii I. Grenberg ◽

Irina A. Grigorieva ◽

...

Keyword(s):

Fourier Transform ◽

20Th Century ◽

Fourier Transform Infrared ◽

Oil Paintings ◽

Ft Ir ◽

Ir Microspectroscopy

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Assessment of material blending distribution for electrospun nanofiber membrane by Fourier transform infrared (FT-IR) microspectroscopy and image cluster analysis

Infrared Physics & Technology ◽

10.1016/j.infrared.2014.06.004 ◽

2014 ◽

Vol 66 ◽

pp. 141-145 ◽

Cited By ~ 2

Author(s):

Jackapon Sunthornvarabhas ◽

Kanjana Thumanu ◽

Wanwisa Limpirat ◽

Hyun-Joong Kim ◽

Kuakoon Piyachomkwan ◽

...

Keyword(s):

Cluster Analysis ◽

Fourier Transform ◽

Fourier Transform Infrared ◽

Electrospun Nanofiber ◽

Nanofiber Membrane ◽

Ft Ir ◽

Ir Microspectroscopy

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High-Throughput Analyses of Microplastic Samples Using Fourier Transform Infrared and Raman Spectrometry

Applied Spectroscopy ◽

10.1177/0003702820932926 ◽

2020 ◽

Vol 74 (9) ◽

pp. 1185-1197 ◽

Cited By ~ 2

Author(s):

Josef Brandt ◽

Lars Bittrich ◽

Franziska Fischer ◽

Elisavet Kanaki ◽

Alexander Tagg ◽

...

Keyword(s):

Fourier Transform ◽

Open Source Software ◽

Fourier Transform Infrared ◽

Particle Analysis ◽

Particle Detection ◽

Raman Spectrometry ◽

Open Source Software Package ◽

Ft Ir ◽

Ir Microspectroscopy ◽

Ir And Raman

Determining microplastics in environmental samples quickly and reliably is a challenging task. With a largely automated combination of optical particle analysis, Fourier transform infrared (FT-IR), and Raman microscopy along with spectral database search, particle sizes, particle size distributions, and the type of polymer including particle color can be determined. We present a self-developed, open-source software package for realizing a particle analysis approach with both Raman and FT-IR microspectroscopy. Our software GEPARD (Gepard Enabled PARticle Detection) allows for acquiring an optical image, then detects particles and uses this information to steer the spectroscopic measurement. This ultimately results in a multitude of possibilities for efficiently reviewing, correcting, and reporting all obtained results.

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Fourier-Transform Infrared Microspectroscopy, a Novel and Rapid Tool for Identification of Yeasts

Applied and Environmental Microbiology ◽

10.1128/aem.68.10.4717-4721.2002 ◽

2002 ◽

Vol 68 (10) ◽

pp. 4717-4721 ◽

Cited By ~ 101

Author(s):

Mareike Wenning ◽

Herbert Seiler ◽

Siegfried Scherer

Keyword(s):

Fourier Transform ◽

Agar Plate ◽

Debaryomyces Hansenii ◽

Fourier Transform Infrared ◽

Strain Level ◽

Correct Identification ◽

Ft Ir ◽

Small Model ◽

Ir Microspectroscopy ◽

Rapid Tool

ABSTRACT Fourier-transform infrared (FT-IR) microspectroscopy was used in this study to identify yeasts. Cells were grown to microcolonies of 70 to 250 μm in diameter and transferred from the agar plate by replica stamping to an IR-transparent ZnSe carrier. IR spectra of the replicas on the carrier were recorded using an IR microscope coupled to an IR spectrometer, and identification was performed by comparison to reference spectra. The method was tested by using small model libraries comprising reference spectra of 45 strains from 9 genera and 13 species, recorded with both FT-IR microspectroscopy and FT-IR macrospectroscopy. The results show that identification by FT-IR microspectroscopy is equivalent to that achieved by FT-IR macrospectroscopy but the time-consuming isolation of the organisms prior to identification is not necessary. Therefore, this method also provides a rapid tool to analyze mixed populations. Furthermore, identification of 21 Debaryomyces hansenii and 9 Saccharomyces cerevisiae strains resulted in 92% correct identification at the strain level for S. cerevisiae and 91% for D. hansenii, which demonstrates that the resolution power of FT-IR microspectroscopy may also be used for yeast typing at the strain level.

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Optimization of Sample Preparation Using Glass Slides for Spectral Pathology

Applied Spectroscopy ◽

10.1177/0003702820945748 ◽

2020 ◽

pp. 000370282094574

Author(s):

Lewis M. Dowling ◽

Paul Roach ◽

Abigail V. Rutter ◽

Ibraheem Yousef ◽

Srinivas Pillai ◽

...

Keyword(s):

Lung Cancer ◽

Sample Preparation ◽

Cancer Cells ◽

Lung Cancer Cells ◽

Thin Glass ◽

Glass Substrates ◽

Standard Clinical Practice ◽

Different Types ◽

Ft Ir ◽

Ir Microspectroscopy

The clinical translation of Fourier transform infrared (FT-IR) microspectroscopy in pathology will require bringing this technique as close as possible to standard practice in pathology departments. An important step is sample preparation for both FT-IR microspectroscopy and pathology. This should entail minimal disruption of standard clinical practice while achieving good quality FT-IR spectral data. In fact, the recently described possibility of obtaining FT-IR spectra of cells placed on glass substrates brings FT-IR microspectroscopy closer to a clinical application. We have now furthered this work in order to identify two different types of lung cancer cells placed on glass coverslips. Two types of sample preparation which are widely used in pathology, cytospin and smear, have been used. Samples were fixed with either methanol, used in pathology, or formalin (4% paraformaldehyde) used widely in spectroscopy. Fixation with methanol (alcohol-based fixative) removed lipids from cells causing a decrease in intensity of the peaks at 2850 cm−1 and 2920 cm−1. Nevertheless, we show for the first time that using either type of sample preparation and fixation on thin glass coverslips allowed to differentiate between two different types of lung cancer cells using either the lipid region or the fingerprint region ranging from 1800 cm−1 to 1350 cm−1. We believe that formalin-fixed cytospin samples would be preferred to study cells on thin coverslips using FT-IR microspectroscopy. This work presents a clear indication for future advances in clinical assessment of samples within pathology units to gain a deeper understanding of cells/tissues under investigation.

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