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Holotomography-driven learning unlocks in-silico staining of single cells in flow cytometry by avoiding fluorescence co-registration

Title: Holotomography-driven learning unlocks in-silico staining of single cells in flow cytometry by avoiding fluorescence co-registration
Authors: Pirone, Daniele; Giugliano, Giusy; Schiavo, Michela; Montella, Annalaura; Mugnano, Martina; Cerbone, Vincenza; Raia, Maddalena; Scalia, Giulia; Kurelac, Ivana; Medina, Diego Luis; Miccio, Lisa; Capasso, Mario; Iolascon, Achille; Memmolo, Pasquale; Ferraro, Pietro
Contributors: Pirone, Daniele; Giugliano, Giusy; Schiavo, Michela; Montella, Annalaura; Mugnano, Martina; Cerbone, Vincenza; Raia, Maddalena; Scalia, Giulia; Kurelac, Ivana; Medina, Diego Lui; Miccio, Lisa; Capasso, Mario; Iolascon, Achille; Memmolo, Pasquale; Ferraro, Pietro
Publication Year: 2026
Collection: IRIS Università degli Studi di Bologna (CRIS - Current Research Information System)
Subject Terms: quantitative phase imaging holographic tomography flow cytometry label-free computational microscopy in-silico staining deep learning
Description: Virtual staining is the current state-of-the-art computational technique to cleverly enhance intracellular specificity in unstained biological samples by using convolutional neural networks (CNNs) trained on co-registered pairs of unstained/stained images. While effective, this approach suffers from unpredictable biases inherent to fluorescence microscopy and encounters challenges when applied to flow cytometry data as it would require accurate co-registration on a huge number of images. Here, we present a novel method that exploits for the first time a Holotomography-driven learning to completely eliminate the need for co-registration. We demonstrate that training a CNN on a stain-free dataset of 3D refractive index tomograms of flowing cells unlocks stain-free intracellular specificity for the first time in quantitative phase imaging flow cytometry. This self-supervised solution, by circumventing the critical obstacle of fluorescence co-registration, opens unprecedented perspectives for label-free, high-throughput imaging flow cytometry, offering a powerful new paradigm for advanced 2D and 3D single-cell analysis.
Document Type: article in journal/newspaper
File Description: ELETTRONICO
Language: English
Relation: info:eu-repo/semantics/altIdentifier/wos/WOS:001706400300001; volume:5; issue:2; firstpage:1; lastpage:15; numberofpages:15; journal:Opto-electronic science (Online); https://hdl.handle.net/11585/1049932
DOI: 10.29026/oes.2026.260003
Availability: https://hdl.handle.net/11585/1049932; https://doi.org/10.29026/oes.2026.260003; https://www.oejournal.org/oes/article/doi/10.29026/oes.2026.260003
Rights: info:eu-repo/semantics/openAccess ; license:Licenza per Accesso Aperto. Creative Commons Attribuzione (CCBY) ; license uri:iris.PUB15
Accession Number: edsbas.CF0BDCD0
Database: BASE