Transmission Fourier Transform Infrared Spectroscopic Imaging, Mapping, and Synchrotron Scanning Microscopy with Zinc Sulfide Hemispheres on Living Mammalian Cells at Sub-Cellular Resolution

Ka Lung Andrew Chan; Ali Ibrahim Altharawi; Pedro   Fale; Cai Li  Song; Sergei G.   Kazarian; Gianfelice   Cinque; Valérie Untereiner; Ganesh D  Sockalingum

doi:10.1177/0003702819898275

Transmission Fourier Transform Infrared Spectroscopic Imaging, Mapping, and Synchrotron Scanning Microscopy with Zinc Sulfide Hemispheres on Living Mammalian Cells at Sub-Cellular Resolution

Ka Lung Andrew Chan, Ali Ibrahim Altharawi, Pedro Fale, Cai Li Song, Sergei G. Kazarian, Gianfelice Cinque, Valérie Untereiner, Ganesh D Sockalingum

Institute of Pharmaceutical Science

Research output: Contribution to journal › Article › peer-review

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Abstract

Fourier transform infrared (FT-IR) spectroscopic imaging and microscopy of single living cells are established label-free technique for the study of cell biology. The constant driver to improve the spatial resolution of the technique is due to the diffraction limit given by infrared (IR) wavelength making subcellular study challenging. Recently, we have reported, with the use of a prototype zinc sulfide (ZnS) transmission cell made of two hemispheres, that the spatial resolution is improved by the factor of the refractive index of ZnS, achieving a λ/2.7 spatial resolution using the synchrotron–IR microscopy with a 36× objective with numerical aperture of 0.5. To refine and to demonstrate that the ZnS hemisphere transmission device can be translated to standard bench-top FT-IR imaging systems, we have, in this work, modified the device to achieve a more precise path length, which has improved the spectral quality of the living cells, and showed for the first time that the device can be applied to study live cells with three different bench-top FT-IR imaging systems. We applied focal plane array (FPA) imaging, linear array, and a synchrotron radiation single-point scanning method and demonstrated that in all cases, subcellular details of individual living cells can be obtained. Results have shown that imaging with the FPA detector can measure the largest area in a given time, while measurements from the scanning methods produced a smoother image. Synchrotron radiation single-point mapping produced the best quality image and has the flexibility to introduce over sampling to produce images of cells with great details, but it is time consuming in scanning mode. In summary, this work has demonstrated that the ZnS hemispheres can be applied in all three spectroscopic approaches to improve the spatial resolution without any modification to the existing microscopes.

Original language	English
Pages (from-to)	544-552
Number of pages	9
Journal	APPLIED SPECTROSCOPY
Volume	74
Issue number	5
Early online date	7 Feb 2020
DOIs	https://doi.org/10.1177/0003702819898275
Publication status	Published - 1 May 2020

Keywords

FT-IR imaging
Fourier transform infrared imaging
label-free
lipid
live cell
nucleus
subcellular
zinc sulfide hemispheres spatial resolution

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10.1177/0003702819898275

Transmission Fourier Transform Infrared_CHAN_Acc3Dec2019_GREEN AAMAccepted author manuscript, 1.36 MB

Cite this

Chan, K. L. A., Altharawi, A. I., Fale, P., Song, C. L., Kazarian, S. G., Cinque, G., Untereiner, V., & Sockalingum, G. D. (2020). Transmission Fourier Transform Infrared Spectroscopic Imaging, Mapping, and Synchrotron Scanning Microscopy with Zinc Sulfide Hemispheres on Living Mammalian Cells at Sub-Cellular Resolution. APPLIED SPECTROSCOPY, 74(5), 544-552. https://doi.org/10.1177/0003702819898275

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title = "Transmission Fourier Transform Infrared Spectroscopic Imaging, Mapping, and Synchrotron Scanning Microscopy with Zinc Sulfide Hemispheres on Living Mammalian Cells at Sub-Cellular Resolution",

abstract = "Fourier transform infrared (FT-IR) spectroscopic imaging and microscopy of single living cells are established label-free technique for the study of cell biology. The constant driver to improve the spatial resolution of the technique is due to the diffraction limit given by infrared (IR) wavelength making subcellular study challenging. Recently, we have reported, with the use of a prototype zinc sulfide (ZnS) transmission cell made of two hemispheres, that the spatial resolution is improved by the factor of the refractive index of ZnS, achieving a λ/2.7 spatial resolution using the synchrotron–IR microscopy with a 36× objective with numerical aperture of 0.5. To refine and to demonstrate that the ZnS hemisphere transmission device can be translated to standard bench-top FT-IR imaging systems, we have, in this work, modified the device to achieve a more precise path length, which has improved the spectral quality of the living cells, and showed for the first time that the device can be applied to study live cells with three different bench-top FT-IR imaging systems. We applied focal plane array (FPA) imaging, linear array, and a synchrotron radiation single-point scanning method and demonstrated that in all cases, subcellular details of individual living cells can be obtained. Results have shown that imaging with the FPA detector can measure the largest area in a given time, while measurements from the scanning methods produced a smoother image. Synchrotron radiation single-point mapping produced the best quality image and has the flexibility to introduce over sampling to produce images of cells with great details, but it is time consuming in scanning mode. In summary, this work has demonstrated that the ZnS hemispheres can be applied in all three spectroscopic approaches to improve the spatial resolution without any modification to the existing microscopes.",

keywords = "FT-IR imaging, Fourier transform infrared imaging, label-free, lipid, live cell, nucleus, subcellular, zinc sulfide hemispheres spatial resolution",

author = "Chan, {Ka Lung Andrew} and Altharawi, {Ali Ibrahim} and Pedro Fale and Song, {Cai Li} and Kazarian, {Sergei G.} and Gianfelice Cinque and Val{\'e}rie Untereiner and Sockalingum, {Ganesh D}",

year = "2020",

month = may,

day = "1",

doi = "10.1177/0003702819898275",

language = "English",

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Chan, KLA , Altharawi, AI, Fale, P, Song, CL, Kazarian, SG, Cinque, G, Untereiner, V & Sockalingum, GD 2020, 'Transmission Fourier Transform Infrared Spectroscopic Imaging, Mapping, and Synchrotron Scanning Microscopy with Zinc Sulfide Hemispheres on Living Mammalian Cells at Sub-Cellular Resolution', APPLIED SPECTROSCOPY, vol. 74, no. 5, pp. 544-552. https://doi.org/10.1177/0003702819898275

Transmission Fourier Transform Infrared Spectroscopic Imaging, Mapping, and Synchrotron Scanning Microscopy with Zinc Sulfide Hemispheres on Living Mammalian Cells at Sub-Cellular Resolution. / Chan, Ka Lung Andrew ; Altharawi, Ali Ibrahim; Fale, Pedro et al.
In: APPLIED SPECTROSCOPY, Vol. 74, No. 5, 01.05.2020, p. 544-552.

Research output: Contribution to journal › Article › peer-review

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T1 - Transmission Fourier Transform Infrared Spectroscopic Imaging, Mapping, and Synchrotron Scanning Microscopy with Zinc Sulfide Hemispheres on Living Mammalian Cells at Sub-Cellular Resolution

AU - Chan, Ka Lung Andrew

AU - Altharawi, Ali Ibrahim

AU - Fale, Pedro

AU - Song, Cai Li

AU - Kazarian, Sergei G.

AU - Cinque, Gianfelice

AU - Untereiner, Valérie

AU - Sockalingum, Ganesh D

PY - 2020/5/1

Y1 - 2020/5/1

N2 - Fourier transform infrared (FT-IR) spectroscopic imaging and microscopy of single living cells are established label-free technique for the study of cell biology. The constant driver to improve the spatial resolution of the technique is due to the diffraction limit given by infrared (IR) wavelength making subcellular study challenging. Recently, we have reported, with the use of a prototype zinc sulfide (ZnS) transmission cell made of two hemispheres, that the spatial resolution is improved by the factor of the refractive index of ZnS, achieving a λ/2.7 spatial resolution using the synchrotron–IR microscopy with a 36× objective with numerical aperture of 0.5. To refine and to demonstrate that the ZnS hemisphere transmission device can be translated to standard bench-top FT-IR imaging systems, we have, in this work, modified the device to achieve a more precise path length, which has improved the spectral quality of the living cells, and showed for the first time that the device can be applied to study live cells with three different bench-top FT-IR imaging systems. We applied focal plane array (FPA) imaging, linear array, and a synchrotron radiation single-point scanning method and demonstrated that in all cases, subcellular details of individual living cells can be obtained. Results have shown that imaging with the FPA detector can measure the largest area in a given time, while measurements from the scanning methods produced a smoother image. Synchrotron radiation single-point mapping produced the best quality image and has the flexibility to introduce over sampling to produce images of cells with great details, but it is time consuming in scanning mode. In summary, this work has demonstrated that the ZnS hemispheres can be applied in all three spectroscopic approaches to improve the spatial resolution without any modification to the existing microscopes.

AB - Fourier transform infrared (FT-IR) spectroscopic imaging and microscopy of single living cells are established label-free technique for the study of cell biology. The constant driver to improve the spatial resolution of the technique is due to the diffraction limit given by infrared (IR) wavelength making subcellular study challenging. Recently, we have reported, with the use of a prototype zinc sulfide (ZnS) transmission cell made of two hemispheres, that the spatial resolution is improved by the factor of the refractive index of ZnS, achieving a λ/2.7 spatial resolution using the synchrotron–IR microscopy with a 36× objective with numerical aperture of 0.5. To refine and to demonstrate that the ZnS hemisphere transmission device can be translated to standard bench-top FT-IR imaging systems, we have, in this work, modified the device to achieve a more precise path length, which has improved the spectral quality of the living cells, and showed for the first time that the device can be applied to study live cells with three different bench-top FT-IR imaging systems. We applied focal plane array (FPA) imaging, linear array, and a synchrotron radiation single-point scanning method and demonstrated that in all cases, subcellular details of individual living cells can be obtained. Results have shown that imaging with the FPA detector can measure the largest area in a given time, while measurements from the scanning methods produced a smoother image. Synchrotron radiation single-point mapping produced the best quality image and has the flexibility to introduce over sampling to produce images of cells with great details, but it is time consuming in scanning mode. In summary, this work has demonstrated that the ZnS hemispheres can be applied in all three spectroscopic approaches to improve the spatial resolution without any modification to the existing microscopes.

KW - FT-IR imaging

KW - Fourier transform infrared imaging

KW - label-free

KW - lipid

KW - live cell

KW - nucleus

KW - subcellular

KW - zinc sulfide hemispheres spatial resolution

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U2 - 10.1177/0003702819898275

DO - 10.1177/0003702819898275

M3 - Article

SN - 0003-7028

VL - 74

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EP - 552

JO - APPLIED SPECTROSCOPY

JF - APPLIED SPECTROSCOPY

IS - 5

ER -

Transmission Fourier Transform Infrared Spectroscopic Imaging, Mapping, and Synchrotron Scanning Microscopy with Zinc Sulfide Hemispheres on Living Mammalian Cells at Sub-Cellular Resolution

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