Label-free Raman hyperspectral imaging of single cells cultured on polymer substrates

While Raman hyper-spectral imaging has been widely used for label-free mapping of biomolecules in cells, these measurements require the cells to be cultured on weakly Raman scattering substrates. However, many applications in biological sciences and engineering require the cells to be cultured on po...

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Main Authors: Sinjab, Faris, Sicilia, Giovanna, Shipp, Dustin, Marlow, Maria, Notingher, Ioan
Format: Article
Published: SAGE 2017
Subjects:
Online Access:https://eprints.nottingham.ac.uk/42781/
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author Sinjab, Faris
Sicilia, Giovanna
Shipp, Dustin
Marlow, Maria
Notingher, Ioan
author_facet Sinjab, Faris
Sicilia, Giovanna
Shipp, Dustin
Marlow, Maria
Notingher, Ioan
author_sort Sinjab, Faris
building Nottingham Research Data Repository
collection Online Access
description While Raman hyper-spectral imaging has been widely used for label-free mapping of biomolecules in cells, these measurements require the cells to be cultured on weakly Raman scattering substrates. However, many applications in biological sciences and engineering require the cells to be cultured on polymer substrates that often generate large Raman scattering signals. Here, we discuss the theoretical limits of the signal-to-noise ratio in the Raman spectra of cells in the presence of polymer signals and how optical aberrations may affect these measurements. We show that Raman spectra of cells cultured on polymer substrates can be obtained using automatic subtraction of the polymer signals and demonstrate the capabilities of these methods in two important applications: tissue engineering and in-vitro toxicology screening of drugs. Apart from their scientific and technological importance, these applications are examples of the two most common measurement configurations: 1) cells cultured on an optically thick polymer substrate measured using an immersion/dipping objective; 2) cells cultured on a transparent polymer substrate and measured using an inverted optical microscope. In these examples we show that Raman hyperspectral datasets with sufficient quality can be successfully acquired to map the distribution of common biomolecules in cells, such as nucleic acids, proteins and lipids, as well as detecting the early stages of apoptosis. We also discuss strategies for further improvements that could expand the application of Raman hyperspectral imaging on polymer substrates even further in biomedical sciences and engineering.
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spelling nottingham-427812020-05-04T19:20:52Z https://eprints.nottingham.ac.uk/42781/ Label-free Raman hyperspectral imaging of single cells cultured on polymer substrates Sinjab, Faris Sicilia, Giovanna Shipp, Dustin Marlow, Maria Notingher, Ioan While Raman hyper-spectral imaging has been widely used for label-free mapping of biomolecules in cells, these measurements require the cells to be cultured on weakly Raman scattering substrates. However, many applications in biological sciences and engineering require the cells to be cultured on polymer substrates that often generate large Raman scattering signals. Here, we discuss the theoretical limits of the signal-to-noise ratio in the Raman spectra of cells in the presence of polymer signals and how optical aberrations may affect these measurements. We show that Raman spectra of cells cultured on polymer substrates can be obtained using automatic subtraction of the polymer signals and demonstrate the capabilities of these methods in two important applications: tissue engineering and in-vitro toxicology screening of drugs. Apart from their scientific and technological importance, these applications are examples of the two most common measurement configurations: 1) cells cultured on an optically thick polymer substrate measured using an immersion/dipping objective; 2) cells cultured on a transparent polymer substrate and measured using an inverted optical microscope. In these examples we show that Raman hyperspectral datasets with sufficient quality can be successfully acquired to map the distribution of common biomolecules in cells, such as nucleic acids, proteins and lipids, as well as detecting the early stages of apoptosis. We also discuss strategies for further improvements that could expand the application of Raman hyperspectral imaging on polymer substrates even further in biomedical sciences and engineering. SAGE 2017-12-01 Article PeerReviewed Sinjab, Faris, Sicilia, Giovanna, Shipp, Dustin, Marlow, Maria and Notingher, Ioan (2017) Label-free Raman hyperspectral imaging of single cells cultured on polymer substrates. Applied Spectroscopy, 71 (12). pp. 2595-2607. ISSN 1943-3530 Ionizing radiation; Cytokine; Immunity; Cancer http://journals.sagepub.com/doi/10.1177/0003702817715042 doi:10.1177/0003702817715042 doi:10.1177/0003702817715042
spellingShingle Ionizing radiation; Cytokine; Immunity; Cancer
Sinjab, Faris
Sicilia, Giovanna
Shipp, Dustin
Marlow, Maria
Notingher, Ioan
Label-free Raman hyperspectral imaging of single cells cultured on polymer substrates
title Label-free Raman hyperspectral imaging of single cells cultured on polymer substrates
title_full Label-free Raman hyperspectral imaging of single cells cultured on polymer substrates
title_fullStr Label-free Raman hyperspectral imaging of single cells cultured on polymer substrates
title_full_unstemmed Label-free Raman hyperspectral imaging of single cells cultured on polymer substrates
title_short Label-free Raman hyperspectral imaging of single cells cultured on polymer substrates
title_sort label-free raman hyperspectral imaging of single cells cultured on polymer substrates
topic Ionizing radiation; Cytokine; Immunity; Cancer
url https://eprints.nottingham.ac.uk/42781/
https://eprints.nottingham.ac.uk/42781/
https://eprints.nottingham.ac.uk/42781/