Cyclic Immunofluorescence (CycIF) & Spatial Biology Services

High-plex multiplex immunofluorescence and spatial biology workflows for tumor microenvironment analysis, immune profiling, biomarker discovery, and translational research powered by Lunaphore COMET™ technology.

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Lunaphore COMET™ Multiplex Tissue Imaging

Automated cyclic immunofluorescence workflows for scalable high-dimensional tissue imaging and spatial biomarker analysis.

What is Cyclic Immunofluorescence (CyCIF)?

CyCIF enables repeated cycles of antibody staining and fluorescence imaging to visualize dozens of protein biomarkers within intact tissue architecture while preserving spatial context and cellular organization.

Multiplex Immunofluorescence & Spatial Biology Services:

IMCO provides end-to-end CyCIF services including experimental design consultation, antibody panel optimization, tissue imaging, spatial analysis, and downstream bioinformatics support.

Why we use COMET cycIF in IMCO labs:

  • Ultra-high plexing: simultaneous visualization of 30+ markers in a single tissue section with precise spatial resolution.

  • Same slide H&E for AI driven data analysis.

  • Spatial biology, uncoupled from single-marker limitations: understand cell–cell interactions, neighborhoods, and microenvironment patterns.

  • FFPE-compatible workflows: work with routinely collected clinical samples without complex preprocessing.

  • Rapid, automated workflow: streamlined protocols reduce hands-on time and improve consistency.

  • Quantitative, reproducible data: standardized cycles enable robust image analysis and cross-sample comparability.

  • End-to-end readiness: integrates with our imaging platforms and data analytics to accelerate discovery and decision-making. Learn more about the Lunaphore Comet here.

Flowchart of pathology tissue preparation and imaging process, including steps for antigen retrieval, antibody loading, automated sequencing, image preprocessing, and analysis, with examples of image export and downstream applications.
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Featured Panels

A blue slide displaying a list of 13 names of core biological markers or proteins, including a-SMA, CD11c, Foxp3, CCR2, CD20, GRZB, CD3, CD45, Ki-67, CD8, CD68, and PANCK.

Core Panel

Targets: CD8 T Cells, Tregs, Th Cells, B Cells, Monocytes, Macrophages, DCs, Neoplastic Cells, Fibroblast-like Cells

Function: Proliferation, Cytotoxicity

An orange discovery panel listing 20 items related to cellular biology, including markers like a-SMA, CD8, CD45, CD163, Ki-67, CCR2, CD11b, CD68, Foxp3, PANCK, CD3, CD11c, CD69, GRZB, PD-1, CD4, CD20, CD103, HLA-II, PD-L1.

Discovery Panel

Targets: CD8 T Cells, Tregs, Th Cells, Unconventional T Cells, B Cells, Monocytes, Macrophages, DCs, Other APCs, Neoplastic Cells, Fibroblast-like Cells

Function: Proliferation, Activation, Checkpoint, Cytotoxicity, Tissue Residency, Anti-inflammatory(M2-like)

Green chart titled "Discovery T Function (29)" listing various cell markers and proteins such as CD11c, CD68, GRZB, PD-L1, with rows of other markers and proteins including CD3, CD4, CD8, and CD45.

Discovery T-Function

Targets: CD8 T Cells, T-regs, Th1 Cells, Th2 Cells, Unconventional Cells, B Cells, Monocytes, Macrophages, DCs, Other APCs, Neoplastic Cells, Fibroblast-like Cells

Function: Proliferation, Activation, Cytotoxicity, Tissue Residency, Memory, Exhaustion, Self-renewal, Immunosuppression, Anti-inflammatory (M2-like)

Custom Panels

Chart titled 'AML Bone Marrow (19)' listing various cell markers, including a-SMA, CD11c, CD45RA, CD163, CD3, CD20, CD61, Foxp3, CD4, CD31, CD68, Ki-67, CD8, CD34, CD71, MPO, CD11b, CD45, CD117, with some markers highlighted in orange.
A table listing 16 different cell markers related to diffuse large B-cell lymphoma (DLBCL), including CD3, CD20, EOMES, LAG-3, CD4, CD45, FOXP3, PD-1, CD8, CD45RA, GRZB, TBET, CD19, CD69, Ki-67, and TIM-3.
Table showing neural cell markers, with some in white and others in orange, titled 'Neuronal (13)' on a light blue background.
A presentation slide titled "Carcinoma (20)" listing various immunohistochemical markers such as a-SMA, CD11b, CD69, HLA-II, CCR2, CD11c, CD103, Ki-67, CD3, CD20, CD163, PANCK, CD4, CD45, Foxp3, PD-1, CD8, CD68, GRZB, PD-L1.
A table titled 'Melanoma (17)' listing various biomarkers and genes associated with melanoma, including Melanosome and SOX-10 in orange text.
Table titled 'Dermatofibrosarcoma Protuberans (DFSP) (32)'. It lists various cell surface markers, with some listed in white, and CD21 and CD23 in orange. The markers include a-SMA, CD20, CD68, GATA-3, PD-1, CCR2, CD69, GRZB, PD-L1, CD3, CD103, HLA-II, TBET, CD4, CD39, CD163, Ki-67, TCF1, CD8, CD45, DC-LAMP, LAG-3, TIM-3, CD11b, CD45RA, Foxp3, PANCK, TOX, CD11c, CD45RO.
A chart titled 'Melanoma - Fresh Frozen (33)' listing various genetic markers and proteins associated with melanoma, including CD20, CD39, PD-1, and BRAF, with some in orange and others in blue.

Lunaphore at work

  • A digital, neon-like abstract art piece featuring a human face outline in orange lines intertwined with blue digital particles on a black background.

    Melanoma: DAPI, POSTN

  • Close-up of cells stained with fluorescent antibodies, showing green fluorescence on the left and multicolored fluorescence on the right, used for scientific research.

    Sarcoma: CD3, CD8, Pan-CK

  • Breast Cancer: Pan-CK, CD45

    Breast Cancer: Pan-CK, CD45

  • Breast Cancer: Pan-CK, CD45

    Breast Cancer: Pan-CK, CD45

  • Melanoma TLS: CD8, ASMA

    Melanoma TLS: CD8, ASMA

  • Lung: CD4, CD8, ASMA, Pan-CK

    Lung: CD4, CD8, ASMA, Pan-CK

  • Melanoma TLS: Ki-67

    Melanoma TLS: Ki-67

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FAQs

  • Cyclic immunofluorescence (CyCIF) is a high-plex imaging technique that uses repeated cycles of antibody staining, fluorescence imaging, and signal removal to visualize dozens of protein biomarkers within a single tissue section. This approach preserves tissue architecture and spatial context, enabling comprehensive characterization of cellular phenotypes and tissue organization.

  • Traditional immunofluorescence is typically limited to a small number of biomarkers due to spectral overlap between fluorophores. CyCIF overcomes this limitation by performing multiple staining and imaging cycles, allowing significantly more biomarkers to be analyzed within the same tissue section.

  • CyCIF is commonly performed on formalin-fixed paraffin-embedded (FFPE) tissue and can also be applied to other validated tissue types depending on study requirements. IMCO works with investigators to determine the most appropriate sample preparation and workflow for each project.

  • CyCIF is widely used for tumor microenvironment characterization, immune profiling, spatial biology, biomarker discovery, translational pathology, neuroinflammation research, and studies requiring high-dimensional tissue imaging while preserving spatial relationships between cells.

  • The Lunaphore COMET™ is a fully automated cyclic immunofluorescence platform that enables high-plex multiplex tissue imaging through iterative staining and imaging cycles. It delivers reproducible, scalable spatial biology workflows for translational research and biomarker discovery.

  • The number of biomarkers depends on the antibody panel design and study objectives. CyCIF supports high-plex imaging workflows capable of measuring dozens of protein biomarkers within a single tissue section while maintaining spatial context. We commonly operate panels of over 30 markers in a single run.

  • Yes. In addition to tissue staining and imaging, IMCO provides downstream spatial analysis services including single-cell segmentation, cell phenotyping, neighborhood analysis, biomarker co-expression analysis, tissue architecture mapping, and publication-ready data visualization.

  • Yes. CyCIF data can be integrated with complementary datasets such as Olink® proteomics, flow cytometry, mass cytometry (CyTOF), genomics, and transcriptomics to provide a more comprehensive understanding of biological systems and disease mechanisms.

  • Unlike conventional IHC, which typically evaluates one or a few markers per tissue section, CyCIF enables simultaneous analysis of dozens of biomarkers while preserving spatial context. This provides deeper insights into cell populations, cellular interactions, and tissue organization.

  • Our team collaborates with investigators throughout the project lifecycle, providing guidance on study design, antibody panel selection, tissue preparation, imaging workflows, quality control, spatial analysis, and biological interpretation to ensure data are aligned with each study's scientific objectives.