Multiplexed Tissue Imaging Platform

Cell DIVE™ is a powerful and innovative commercial technology platform that enables a researcher to analyze single 5 um tissue section with up to 60 biomarker stains. This platform has been developed over 10 years with a technically diverse team and multiple external collaborators. The process involves iteratively staining, imaging and inactivating a single tissue slide with sequential rounds of dye-labeled antibodies. Our multidisciplinary team has developed custom imaging software and algorithms including image registration, quality, cross-microscope calibration and down-stream image processing including image correction and cell segmentation. 

As we developed this platform and understood our customer and collaborator needs, we focused on microscope calibration as an essential component of data quality and the development of a common reference standard. Our approach is to use reference values or idealized models for each calibration parameter and calibrate each microscope relative to these parameters. The reference values do not change and therefore each microscope can be considered independently and calibrated independently to the standard. Periodic checks or routine automated calibrations can then be used to determine if any of the calibration parameters have deviated too far from their calibrated values, and recalibrate if necessary.  This robust calibration scheme was accomplished through the use of custom calibration plates that contained the appropriate targets. Mechanical, optical, and geometric calibrations were developed using the calibration plate (as the target), custom scripts and custom algorithms. 

Today, our Cell DIVE platforms and workflows are available as a service (MultiOmyx™ with Neogenomics) and are increasingly sought after by research labs, as interest in multiplexing and multi-marker and single cell analysis grows.