GeoMx ROI Selection Markers

Supplemental Morphology Markers for NanoString® GeoMx® Digital Spatial Profiling Assays

New markers augment off-the-shelf kits and provide further sample stratification aiding in ROI selection of biologically relevant tissue.

Canopy Biosciences ROI Selection Markers for GeoMx® Digital Spatial Profiling

NanoString® GeoMx® DSP assays are a highly sensitive method for the detection of protein and RNA targets in tissue with spatial resolution. Morphology markers elucidate key structures in tissue samples and enable ROI-guided analysis for high-plex transcriptomic and proteomic profiling. Canopy’s ROI Selection Markers enhance analysis by expanding available markers for ROI selection into cell- and disease-specific areas. This allows for further stratification of tissue and selection of biologically relevant samples for downstream profiling.

Customize Your Spatial Transcriptomic Assay

Identifying changes in tumor and tumor microenvironment is central to NanoString® GeoMx® DSP assays. Morphology marker staining reveals tissue architecture and offers insights into selecting the appropriate regions of interest, making the analysis more meaningful. But markers such as PanCK and CD45 only tell part of the story, broadly identifying solid tumors. Canopy’s portfolio of markers allows researchers to add in an additional marker more in line with their study — deciphering between cell types or in a disease-specific manner.

The portfolio consists of markers important to immune profiling and immuno-oncology, and is continuously expanding into application areas relevant to today’s scientists – immuno-oncology, immunology, neuroscience and oncology research.

Marker Identification and Validation

GeoMx® DSP is a fluorescent imaging system that uses standard filter sets FITC, Cy3, Texas Red and Cy5 channels. Antibodies are conjugated to a fluorophore compatible with one of these filter sets to visualize it in the tissue sample. Off the shelf kits from NanoString(R) only use 3 of these channels, leaving one open for customization, so Canopy identified antibodies conjugated to AF647, to coincide with the Cy5 channel. We followed the verification guidelines found in the NanoString(R) Morphology Marker Guidelines document to ensure proper validation for our markers. Additional multiplex validation steps were done to for final antibody confirmation and inclusion in this portfolio.

CD3 marker validated by Canopy Biosciences in tonsil tissue for use in GeoMx DSP ROI selection. This marker visualizes CD3 in tissue samples, a key T cell helper with a critical role in T cell-mediated responses.
CD68 marker validated by Canopy Biosciences in tonsil tissue for use in GeoMx DSP ROI selection. This marker visualizes CD68, a key monocyte and machrophage marker, with a critical role in apoptosis and phagocytosis.
TIGIT marker validated by Canopy Biosciences in tonsil tissue for use in GeoMx DSP ROI selection. This marker highlights TIGIT, a cell surface inhibitory receptor with influence over T cell activation and interleukin production.

Canopy’s knowledge of IHC and histopathology from the Canopy Core Lab provides significant expertise to validate this catalog of markers, and to continue to grow the offering. In addition, for those who need fully custom markers — replacing the other markers in the off-the-shelf kits — Canopy has the capabilities to design and validate completely custom marker sets.

Overview of GeoMx® DSP Workflow

Comprehensive GeoMx® Services

Canopy Biosciences is a full service GeoMx® assay provider. We run both protein and RNA assays for nCounter® and NGS read-outs with the GeoMx® workflow. Our comprehensive service includes live ROI selection with clients and an overview of the Analysis suite for GeoMx® data analysis. Our goal is for our clients to get the most out of the data generated using this platform so we provide training on the analysis software so clients can dig into the wide breadth of data. Our ROI selection markers are just another way we are focusing on helping clients make the most out of these exciting spatial biology experiments.

ROI Selection Marker Portfolio

Canopy’s ROI Selection Marker portfolio is continually expanding to add new markers relevant to today’s critical research application areas. Key target areas include immuno-oncology, immunology, neuroscience and core oncological markers. Each marker further is categorized into aid in selection from our extensive list.

Immune Cell Profiling

Critical markers to identify immune cell types, including T cells, B cells, macrophages, and NK cells. Additional markers for deeper profiling of immune cells, including subtyping T cells.

Immune Cell Activation Status

Critical checkpoint molecules that modulate T cell activation. Key markers of T cell activation mediate the progression of immune response.

Immuno-oncology Drug Targets

Critical drug targets in development with the immuno-oncology field, including many immune checkpoint molecules. Drug targets have the potential to enhance anti-cancer immune responses.

Immune Cell ProfilingImmune Cell ActivationImmuno-oncology Drug Targets
CD3CD27B7-H4
CD4CD28CTLA-4
CD11cOX40IDO
CD8PD-L1LAG-3
CD68OX40L
FoxP3PD-1
Granzyme BTIGIT
TIM-3

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