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

Quantitative phase imaging 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, quantitative phase imaging does not provide the labelling of various specific intracellular structures. Here we show...

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Bibliographic Details
Published in:Nature photonics 2022-12, Vol.16 (12), p.851-859
Main Authors: 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
Format: Article
Language:English
Subjects:
132/124
631/1647/328/1651
631/553/2706
639/624/1107/328/1651
Applied and Technical Physics
Computer applications
Flow cytometry
Fluorescence
Fluorescence microscopy
Fluorometers
Image segmentation
Imaging techniques
Intracellular
Labeling
Medical imaging
Microfluidics
Microscopy
Nuclei
Nuclei (cytology)
Phase contrast
Physics
Physics and Astronomy
Quantum Physics
Refractivity
Staining
Stains
Stains & staining
Statistical inference
Tumor cell lines
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Summary:Quantitative phase imaging 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, quantitative phase imaging does not provide the labelling of various specific intracellular structures. Here we show a novel computational segmentation method based on statistical inference that makes it possible for quantitative phase imaging techniques to identify the cell nucleus. We demonstrate the approach with refractive index tomograms of stain-free cells reconstructed using tomographic phase microscopy in the 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 data and microfluidic cyto-fluorimeter outputs. This is a remarkable step towards directly extracting specific three-dimensional intracellular structures from the phase contrast data in a typical flow cytometry configuration. The accurate identification of the three-dimensional quantitative shape of a cell nucleus is now possible without fluorescent staining by applying computational segmentation to refractive index tomograms recorded in the flow cytometry mode.
ISSN:1749-4885
1749-4893
DOI:10.1038/s41566-022-01096-7