{
  "abstract": "In the present work, we developed a data-driven algorithm for the automated segmentation of fluorescence lifetime imaging microscopy (FLIM) images, enhancing the analysis of multifunctionalized nanoparticles (NPs) within living cancer cells. FLIM, a powerful microscopy technique, generates images that capture the fluorescence lifetime across a sample at the pixel level, revealing critical details about the molecular environment. Traditionally, FLIM image analysis has relied on manual segmentation with the phasor plot approach, a graphical representation of FLIM data in a G and S coordinate system, which is susceptible to user bias and inconsistency. Here, we present an automated and free of user-biased thresholding and segmentation algorithm that streamlines with clustering techniques to automatically identify phasor-clusters in the phasor plot space, reducing user dependency and providing a reproducible strategy under tested conditions image segmentation. We demonstrate its application in the context of FLIM images displaying a map of intensity heterogeneity where functionalized NPs affect the cellular metabolism of HeLa cells, reported by NADH, a bright and a dim fluorescent source, respectively, both of biological relevance. This algorithm provides a transparent and reproducible approach for FLIM image analysis, showing good agreement with expert-defined segmentation under sufficient contrast conditions, while presenting limitations in low-contrast or noisy regimes.",
  "abstract_shown": true,
  "affiliations": [
    "Facultad de Ingeniería, Arquitectura y Diseño, Universidad Autónoma de Baja California, Ensenada, Baja California 22860, Mexico",
    "These authors contributed equally to this work.",
    "Department of Biomedical Engineering, University of California Irvine, Irvine, CA 92697, United States of America",
    "Centro de Nanociencias y Nanotecnología, Ensenada, Baja California 22860, Mexico"
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 5,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Dora-Luz Flores",
      "orcid": "0000-0002-2623-3772"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Esperanza Guerra-Rosas",
      "orcid": "0000-0002-2006-8489"
    },
    {
      "affs": [
        2,
        1
      ],
      "name": "Balam Benítez-Mata",
      "orcid": "0000-0001-8698-8694"
    },
    {
      "affs": [
        3
      ],
      "name": "Gustavo Hirata",
      "orcid": "0000-0002-5120-1027"
    },
    {
      "affs": [
        2
      ],
      "name": "Michelle Digman",
      "orcid": "0000-0003-4611-7100"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/dcw1v4swcsq93.bib",
  "cited_by": 0,
  "doi": "10.1088/2050-6120/ae7fed",
  "fulltext_url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC13733409/",
  "id": "dcw1v4swcsq93",
  "issns": [
    "2050-6120"
  ],
  "landing_url": "https://doi.org/10.1088/2050-6120/ae7fed",
  "language": "en",
  "licence": "CC-BY-4.0",
  "licence_label": "CC BY 4.0",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "heading",
      "Article"
    ],
    [
      "openalex",
      "1304|Advanced Fluorescence Microscopy Techniques"
    ]
  ],
  "openalex": "W7165364676",
  "pdf_url": "https://iopscience.iop.org/article/10.1088/2050-6120/ae7fed/pdf",
  "pmcid": "PMC13733409",
  "pmid": "42320528",
  "published": "2026-07-01",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/dcw1v4swcsq93.ris",
  "sources": [
    {
      "first_seen": "2026-11-08T12:23:11Z",
      "native_id": "PMC13733409",
      "source": "pmc",
      "url": "https://doi.org/10.1088/2050-6120/ae7fed"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1304|Advanced Fluorescence Microscopy Techniques",
      "subfield": "biophysics"
    }
  ],
  "title": "Unsupervised clustering FLIM-phasor from multifunctionalized nanoparticles in living cancer cells",
  "title_text": "Unsupervised clustering FLIM-phasor from multifunctionalized nanoparticles in living cancer cells",
  "type": "peer-reviewed",
  "updated_at": "2026-11-08T12:23:11Z",
  "url": "https://research.draconitic.com/paper/dcw1v4swcsq93",
  "venue": "Methods and applications in fluorescence",
  "versions": []
}