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Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry

Nature photonics, 2022-12, Vol.16 (12), p.851-859 [Peer Reviewed Journal]

The Author(s) 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. ;ISSN: 1749-4885 ;EISSN: 1749-4893 ;DOI: 10.1038/s41566-022-01096-7 ;PMID: 36451849

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Title:
Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry
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
Subjects: Computer applications ; Flow cytometry ; Fluorescence ; Fluorescence microscopy ; Fluorometers ; Image segmentation ; Imaging techniques ; Intracellular ; Labelling ; Medical imaging ; Microfluidics ; Microscopy ; Nuclei ; Nuclei (cytology) ; Phase contrast ; Refractivity ; Staining ; Stains ; Stains & staining ; Statistical inference ; Tumor cell lines
Is Part Of: Nature photonics, 2022-12, Vol.16 (12), p.851-859
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.
Publisher: England: Nature Publishing Group
Language: English
Identifier: ISSN: 1749-4885
EISSN: 1749-4893
DOI: 10.1038/s41566-022-01096-7
PMID: 36451849
Source: ProQuest Central

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Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry

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