This lab is working to create a standardized, high throughput, and user-friendly workflow for using Multiplexed Ion Beam Imaging by Time of Flight (MIBI-TOF) in basic and translational research to gain insight into how single cell phenotype and tissue structure are functionally-linked in health and disease.

To achieve this, they are validating 100 formalin fixed, paraffin embedded tissue (FFPE) antibodies and optimizing ready-to-use multiplexed staining panels in lyophilized format that will permit storage for at least two years. Protocols and reagents for multiplexed signal amplification of protein and mRNA targets are being further refined, while next generation instrumentation are increasing sample throughput to permit full tissue section imaging of up to 40 proteins in 1 hour. Standardized reagents and more robust instrumentation are accompanied by an automated computational pipeline that utilizes a standard set of segmentation markers and machine learning to accurately identify nuclei and cell borders in any non-neural human tissue. This data is used to cluster single cell events into functionally distinct populations according to morphology, protein expression, and histological distribution. The work of this lab provides a practical, back compatible imaging platform for high throughput multiplexed imaging, and will also accelerate development of other complementary imaging technologies.