NSH Annual Meeting

National Society of Histotechnology


September 8 - 12

Baltimore US

Booth #1017

Conference Information

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4 days of cutting-edge research, expert lead workshops and one-of-a-kind hands-on histology training in developing specialties. Explore the largest histology focused expo featuring top industry vendors previewing equipment, supplies, and services for your lab. Create a network of histology experts that continue to assist you when you return to your lab and help to advance your career.

September 10

Poster Presentation

Sunday, September 10

1:15 PM

Multiplex immunofluorescence (mIF) has accelerated the understanding of the spatial immune context in tissues and boosted the identification of new potential biomarkers and therapeutical targets. Despite its increased application in characterization of the tumor microenvironment (TME), state-of-the art protocols remain technically challenging due to dedicated reagents, lengthy procedures, questionable reproducibility, and poor transferability between different tissue types. 

Here, we present the development and validation of an Immuno-Oncology (IO) Core Panel of 13 clinically relevant biomarkers to enable automated spatial analysis of the immune TME on the COMET™ platform across various tissue types.  

Formalin-fixed paraffin-embedded human tissue sections from tonsil and a 24-core multi-organ tissue microarray (TMA) were stained using the IO Core Panel from Lunaphore on the COMET™ platform by fully automated sequential immunofluorescence (seqIF™), which consists of cycles of staining, imaging, and elution. The 13-plex IO Core panel was developed and validated on tonsil. The sections retrieved from COMET™ after seqIF™, were stained by a histology facility with standard immunohistochemistry assay established for routine pathological diagnosis. All markers demonstrated accurate detection with specific staining, comparable to immunohistochemistry counterparts. Subsequently, the panel was transferred on a TMA including several tumoral and non-tumoral specimens, showing robust performance across all samples. The protocol was optimized to achieve high staining quality for all 13 markers in terms of signal specificity, sensitivity, ratio to background and dynamic range. The repeatability and reproducibility of the seqIF™ assay was verified by day-to-day tests on one platform and tests among multiple platforms, respectively.  

Our study demonstrates a validation workflow that highlights the ease of adoption of multiplex imaging to explore TME composition at single-cell resolution with robust IO Core panel, which is easily transferrable to various cohorts of specimens and provides a toolset for spatial cellular dissection of tissue architecture.