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Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry
Quantitative Phase Imaging (QPI) has gained popularity in bioimaging because it can avoid the need for cell staining, which in some cases is difficult or impossible. However, as a result, QPI does not provide labelling of various specific intracellular structures. Here we show a novel computational...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7613862/ https://www.ncbi.nlm.nih.gov/pubmed/36451849 http://dx.doi.org/10.1038/s41566-022-01096-7 |
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author | Pirone, Daniele Lim, Joowon Merola, Francesco Miccio, Lisa Mugnano, Martina Bianco, Vittorio Cimmino, Flora Visconte, Feliciano Montella, Annalaura Capasso, Mario Iolascon, Achille Memmolo, Pasquale Psaltis, Demetri Ferraro, Pietro |
author_facet | Pirone, Daniele Lim, Joowon Merola, Francesco Miccio, Lisa Mugnano, Martina Bianco, Vittorio Cimmino, Flora Visconte, Feliciano Montella, Annalaura Capasso, Mario Iolascon, Achille Memmolo, Pasquale Psaltis, Demetri Ferraro, Pietro |
author_sort | Pirone, Daniele |
collection | PubMed |
description | Quantitative Phase Imaging (QPI) has gained popularity in bioimaging because it can avoid the need for cell staining, which in some cases is difficult or impossible. However, as a result, QPI does not provide labelling of various specific intracellular structures. Here we show a novel computational segmentation method based on statistical inference that makes it possible for QPI techniques to identify the cell nucleus. We demonstrate the approach with refractive index tomograms of stain-free cells reconstructed through the tomographic phase microscopy in flow cytometry mode. In particular, by means of numerical simulations and two cancer cell lines, we demonstrate that the nucleus can be accurately distinguished within the stain-free tomograms. We show that our experimental results are consistent with confocal fluorescence microscopy (FM) data and microfluidic cytofluorimeter outputs. This is a significant step towards extracting specific three-dimensional intracellular structures directly from the phase-contrast data in a typical flow cytometry configuration. |
format | Online Article Text |
id | pubmed-7613862 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
record_format | MEDLINE/PubMed |
spelling | pubmed-76138622023-05-10 Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry Pirone, Daniele Lim, Joowon Merola, Francesco Miccio, Lisa Mugnano, Martina Bianco, Vittorio Cimmino, Flora Visconte, Feliciano Montella, Annalaura Capasso, Mario Iolascon, Achille Memmolo, Pasquale Psaltis, Demetri Ferraro, Pietro Nat Photonics Article Quantitative Phase Imaging (QPI) has gained popularity in bioimaging because it can avoid the need for cell staining, which in some cases is difficult or impossible. However, as a result, QPI does not provide labelling of various specific intracellular structures. Here we show a novel computational segmentation method based on statistical inference that makes it possible for QPI techniques to identify the cell nucleus. We demonstrate the approach with refractive index tomograms of stain-free cells reconstructed through the tomographic phase microscopy in flow cytometry mode. In particular, by means of numerical simulations and two cancer cell lines, we demonstrate that the nucleus can be accurately distinguished within the stain-free tomograms. We show that our experimental results are consistent with confocal fluorescence microscopy (FM) data and microfluidic cytofluorimeter outputs. This is a significant step towards extracting specific three-dimensional intracellular structures directly from the phase-contrast data in a typical flow cytometry configuration. 2022-12 2022-11-10 /pmc/articles/PMC7613862/ /pubmed/36451849 http://dx.doi.org/10.1038/s41566-022-01096-7 Text en https://www.springernature.com/gp/open-research/policies/accepted-manuscript-termsUsers may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: https://www.springernature.com/gp/open-research/policies/accepted-manuscript-terms |
spellingShingle | Article Pirone, Daniele Lim, Joowon Merola, Francesco Miccio, Lisa Mugnano, Martina Bianco, Vittorio Cimmino, Flora Visconte, Feliciano Montella, Annalaura Capasso, Mario Iolascon, Achille Memmolo, Pasquale Psaltis, Demetri Ferraro, Pietro Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry |
title | Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry |
title_full | Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry |
title_fullStr | Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry |
title_full_unstemmed | Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry |
title_short | Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry |
title_sort | stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7613862/ https://www.ncbi.nlm.nih.gov/pubmed/36451849 http://dx.doi.org/10.1038/s41566-022-01096-7 |
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