Enhancing Multiplex Immunofluorescence Workflows

An Automated End-to-End Spatial Biology Workflow for Improved Efficiency and Reproducibility

ZEISS Microscopy Solutions

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Boosting Speed and Accuracy Through Automation

Background

Spatial biology is evolving rapidly from primary research to routine application, with an increasing focus on workflow efficiency, result accuracy, and reproducibility. Multiplex immunofluorescence (mIF) stands out as a powerful tool to characterize multiparametric disease processes within the tumor microenvironment (TME).

Despite its high potential, implementing mIF protocols for routine research can be challenging on multiple levels: mIF staining protocols are more complex to establish than traditional chromogenic staining protocols and reveal significant variability intrinsic to tissue samples, which needs to be accounted for in the image acquisition and analysis process. The latter challenges are usually addressed by manual image acquisition and analysis processes which are time-consuming and introduce additional sources of variability preventing comparability of results.

To address these problems, we present a fully pre-optimized and validated workflow with high degree of automation, combining the high reproducibility of pre-optimized Ultivue's InSituPlex® (ISP) multiplex immunofluorescent assay technology with a user-friendly automated image acquisition process for the ZEISS Axioscan 7 spatial biology and automated image analysis on the new Mindpeak mIF analysis platform (Figure 1). This approach allows for a highly streamlined workflow which significantly reduces hands on time and provides results with enhanced reproducibility across operators.