Value of multiphoton microscopy in uro-oncology - a narrative review

Sep 24, 2026

Multi-photon microscopy (MPM) is a three-dimensional fluorescence imaging approach that uses low-energy photon excitation to reduce photo-bleaching while enabling deeper tissue penetration. It detects autofluorescence from intrinsic intracellular fluorophores and second harmonic generation to visualize non-centrosymmetric structures such as collagen. A structured literature review using PubMed and Web of Science summarizes how MPM supports biopsy, surgical-margin assessment, staging and grading, and real-time intraoperative imaging, enabling quantitative AF/SHG measurements aligned with clinical outcomes.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages11

Reading Time28 min

AccessFree to download

Introduction

Methods

Key content and findings

Conclusions

Value of multiphoton microscopy in uro-oncology: a narrative review Patrick-Julien Treacy1,2, Akshita Khosla1, Natasha Kyprianou1,3, Ugo Giovanni Falagario1,4, Nikos Tsavaras5, Peter Wiklund1, Ashutosh K. Tewari1, Matthieu Durand2,6 1Department of Urology, Icahn School of Medicine at Mount Sinai, New York, NY, USA; 2Department of Urology, Andrology and Renal Transplantation Unit, Pasteur 2 Hospital, Nice, PACA, France; 3Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA; 4Department of Urology and Organ Transplantation, University of Foggia, Foggia, Puglia, Italy; 5Microscopy Core at Icahn School of Medicine at Mount Sinai Hospital, New York, NY, USA; 6INSERM U1081-CNRS UMR 7284, Nice University Côte d’Azur, Nice, PACA, France Contributions: (I) Conception and design: PJ Treacy, M Durand; (II) Administrative support: AK Tewari; (III) Provision of study materials or patients: A Khosla, N Kyprianou; (IV) Collection and assembly of data: UG Falagario; (V) Data analysis and interpretation: None; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors. Correspondence to: Patrick-Julien Treacy, MD. Department of Urology, Icahn School of Medicine at Mount Sinai, 1425 Madison Avenue, 6th Floor, New York City, NY, 10029, [USA. Email: pj.treacy@live.fr](USA. Email: pj.treacy@live.fr). Background and Objective: Multi-photon microscopy (MPM) is a 3-dimension fluorescence imaging technique that combines the excitation of two low-energy photons, enabling less photo-bleaching and deeper penetration of the imaged tissue. Two signals are detected, autofluorescence (AF), from natural intracellular fluorophores [such as nicotinamide adenine dinucleotide phosphate (NADP) and flavine adenine dinucleotide (FAD) transformation], and second harmonic generation (SHG), a physical property of the laser enhancing non-centrosymmetric structures such as collagen fibers. MPM can give both visual and quantitative information of a fresh tissue (without the need of processing, cutting or staining the tissue), aiding in the progress towards optimizing a real-time imaging device. The objective of this review is to show the value and benefits of the use of MPM in uro-oncology. Methods: A structured literature review was performed using PubMed and Web of Sciences, including all articles with the following keywords: “multiphoton microscopy”, “two-photon microscopy”, “non-linear microscopy”,“second harmonic generation”,“urology”, “prostate”,“bladder”,“kidney”,“upper tract”,“oncology”,“surgical margins”,“frozen section”. Articles were reviewed to summarize the use of this tool in performing biopsies, assessing surgical margins, staging and grading complementary tool, and real-time imaging. Key Content and Findings: A total of 476 articles were identified with these keywords, and later screened for inclusion. We finally included 47 publications that were relevant to our topic. The advantages of this technique have led to its application in the management of several cancers, allowing cellular description as well as quantitative measurements ofAF or SHG and their correlation with clinical outcomes. Conclusions: MPM has shown great improvement in providing a real time assessment of fresh tissue, giving oncologic diagnosis, performing in vivo imaging and quantitative analysis of the tissue as well as increasing precision of the diagnosis. This nonlinear optical technique has the potential of guiding both biopsy and surgery, as well as helping the surgeon with interesting additional tissue information intra operatively. Keywords: Multi-photon microscopy (MPM); connective tissue; in vivo imaging; non-linear microscopy; biopsy Submitted Nov 04, 2021. Accepted for publication Jan 17, 2023. Published online Feb 17, 2023. doi: 10.21037/tau-21-973 View this article at: [https://dx.doi.org/10.21037/tau-21-973](https://dx.doi.org/10.21037/tau-21-973) © Translational Androl

FAQs

What does multiphoton microscopy (MPM) measure in uro-oncology?

MPM detects autofluorescence from intrinsic intracellular fluorophores and second harmonic generation signals from non-centrosymmetric structures such as collagen fibers.

How was the evidence collected for this narrative review?

A structured literature review was conducted in PubMed and Web of Science using specified keywords related to multiphoton/second harmonic imaging and uro-oncology applications; 47 relevant publications were included.

What practical clinical uses does the review highlight for MPM?

The review emphasizes applications in biopsies, assessing surgical margins, aiding staging and grading, and enabling real-time in vivo and intraoperative imaging with quantitative AF/SHG analysis.

Sep 24, 2026

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