Multiplexed Ion Beam Imaging (MIBI) – Methodology and Applications
I will present the MIBI platform, and explain its workflow from the perspective of a core facility.
Characterization of the cellular composition and spatial organization of the tissue microenvironment has been limited by the techniques available to image the necessary number of biomarkers for broad phenotyping at a subcellular scale.
MIBI combines time-of-flight secondary ion mass spectrometry (ToF-SIMS) with metal labeled conventional antibodies to image 40+ biomarkers in situ in a single scan at subcellular spatial resolution.
MIBI starts with a FFPE, fresh, or frozen tissue. The section is stained, following a protocol similar to conventional IHC, with a panel of about 40 antibodies labeled with metal isotopes. Antibodies validation is done via IHC (or IF) before conjugation to an isotope, then the conjugated antibody performance is measured by MIBI and compared to IHC.
MIBI enables quantitative, full periodic table coverage across a five-log dynamic range, imaging all of the labeled antibodies simultaneously, with fields of view up to 800 um × 800 um at resolutions down to 260 nm with sensitivities approaching single antibody detection.
Pre-processing of the images, includes removal of channel crosstalk, noise and antibody aggregates. This is followed by segmentation into single cells, from which frequencies of different cell subsets and the distributions of nearest neighbor distances between them can be calculated.
I will illustrate the platforms’ capabilities through a cohort of clinical tissue sections taken from a case study focusing on characterization of the sentinel lymph nodes microenvironment in relation to melanoma metastasis.
Tomer-Meir Salame received his Ph.D. in Biotechnology, with distinction, from the Hebrew University in 2014. He won over thirty awards and scholarships, including the Wolf fellowship, The Robert H. Smith Vision Prize and the Rector’s award.
Tomer joined the Weizmann Institute of Science, Life Sciences Core Facilities’ Flow Cytometry Unit in 2014 and became a staff scientist in 2017. There he has been collaborating on over 30 multidisciplinary projects, with over 40 peer-reviewed publications and teaching in the cytometry student’s courses. He is also a reviewer at the ISAC SRL Recognition Program Sub-Committee.
Tomer is leading the implementation of mass cytometry via CyTOF and MIBI in research at the Weizmann Institute.
Flow Cytometry Unit, Life Sciences Core Facilities, Weizmann Institute of Science Ullman building, floor 0, room 5, P.O. Box 26, Rehovot 7610001, Israel