{
  "abstract": "Multicolor fluorescence microscopy is central to biological imaging, but simultaneous detection of additional colors typically reduces throughput, increases spectral crosstalk, or lowers signal-to-noise ratio. We developed symmetric off-axis spectral encoding microscopy (sym-OSEM), which uses paired spatial encoding and differential decoding to separate fluorescence channels and suppress common-mode out-of-focus background. The differential decoding requires no specimen-specific reference spectrum, point spread function, or training data. All fluorophores are excited and detected simultaneously, so acquisition time is largely independent of channel count within the demonstrated two- to six-color configurations. Simulations and experiments using cell preparations, a serially imaged 300 μm-thick mouse-brain volume, and lymph node sections validated the approach. Four-color imaging of U2OS cells covered 6.362 mm2 min-1, 59-fold faster than a commercial confocal microscope under the tested acquisition settings, while retaining comparable image quality. Simultaneous six-color imaging resolved the distributions of immune-cell and stromal markers in lymph-node tissue. Sym-OSEM therefore provides a strategy for combining multicolor separation, optical sectioning, and high-throughput acquisition in a single imaging framework.",
  "abstract_shown": true,
  "affiliations": [
    "MoE Key Laboratory for Biomedical Photonics Wuhan National Laboratory for Optoelectronics Huazhong University of Science and Technology Wuhan China."
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 6,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Jiangjiang Zhao",
      "orcid": "0009-0007-9253-0592"
    },
    {
      "affs": [
        0
      ],
      "name": "Zhangheng Ding"
    },
    {
      "affs": [
        0
      ],
      "name": "Zhan Zhang"
    },
    {
      "affs": [
        0
      ],
      "name": "Jie Zhang"
    },
    {
      "affs": [
        0
      ],
      "name": "Shuhong Qi"
    },
    {
      "affs": [
        0
      ],
      "name": "Jing Yuan",
      "orcid": "0000-0001-9050-4496"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/577kqw1vn16y1.bib",
  "cited_by": 0,
  "doi": "10.1002/nap2.70367",
  "fulltext_url": "https://europepmc.org/articles/PMC13744097",
  "id": "577kqw1vn16y1",
  "issns": [
    "2192-8606",
    "2192-8614"
  ],
  "landing_url": "https://europepmc.org/article/MED/42999791",
  "language": "eng",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1304|Advanced Fluorescence Microscopy Techniques"
    ]
  ],
  "openalex": "W7225453818",
  "pdf_url": "https://europepmc.org/articles/PMC13744097?pdf=render",
  "pmcid": "PMC13744097",
  "pmid": "42999791",
  "published": "2026-11-07",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/577kqw1vn16y1.ris",
  "sources": [
    {
      "first_seen": "2026-11-09T15:20:34Z",
      "native_id": "MED:42999791",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42999791"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1304|Advanced Fluorescence Microscopy Techniques",
      "subfield": "biophysics"
    }
  ],
  "title": "Symmetric Off-Axis Spectral Encoding Enables High-Throughput Six-Color Optical Sectioning Imaging.",
  "title_text": "Symmetric Off-Axis Spectral Encoding Enables High-Throughput Six-Color Optical Sectioning Imaging.",
  "type": "peer-reviewed",
  "updated_at": "2026-11-09T15:20:34Z",
  "url": "https://research.draconitic.com/paper/577kqw1vn16y1",
  "venue": "Nanophotonics (Berlin, Germany)",
  "versions": []
}