TRANSFORMING MULTIPLEXED TISSUE IMAGING

MIBIscopeTMWITH MULTIPLEXED ION BEAM IMAGING (MIBI ) TECHNOLOGYTRANSFORMINGMULTIPLEXED TISSUE IMAGING

MIBIscopeFOR ANALYSIS OFTHE TUMOR MICROENVIRONMENTCELL CLASSIFICATIONComprehensivelyphenotype immune infiltrateCOEXPRESSION OF PD-L1 ON VARIOUS CELL TYPESNUMBER OF CELLSCOEXPRESSING pHexictotoCyTllscesgehaproacMBllsgtin sra elllife r Cro onp TumceNoPROTEIN EXPRESSIONQuantify protein expressionPD-L1 EXPRESSION12010080604020MacrophagesDendritic CellsT CellsSPATIAL ANALYSISProfile tissue architectureDISTANCE BETWEEN IMMUNE AND TUMOR CELLSDENSITY OF IMMUNE-TUMORNEAREST NEIGHBORSA REVOLUTIONARY TECHNOLOGY0510152025DISTANCE FROM IMMUNE TO TUMOR CELL (µm)30

C IMHC IICD31iNOSHH3H3K9acFOXP3COL1CD206Na/K 62CD68CD103CD36CD116CARBONVISUALIZE 40 MARKERSIN A SINGLE IMAGE250 µmLung granuloma (3x3 mm2)CD45CD31SMACD6850 µmCD45 CD31 SMA CD68 HH3CD163 CD11c CD8 CD3 HH3

HIGH RESOLUTIONHIGH THROUGHPUTAchieve confocal resolutionImage up to 90 800x800 µm2 ROIs per dayHIGH SENSITIVITYDETECTION 0Pr5Nd5Ce4La310Y215InCytokeratinCD451DNAAb COPY NUMBER2010 ACD4FOXP3DNAData adapted from Keren et al.,Science Advances, 2019

PUBLICATION QUALITYDATAUNMATCHEDREPRODUCIBILITY108dsDNA 89YCOUNTS[ R 0.05 P 0.7 ]Analyzelarge clinicalcohorts106b-catenin 166Er[ R 0.09 P 0.6 ]CD56 145Nd104[ R 0.007 P 0.9 ]102100051015ACQUISITION TIME[DAYS]The MIBIscope has minimal signal drift overmultiple weeks of data collection, so largeclinical trials can be imaged and analyzedwith confidence.Triple negative breast cancerH3K27me3b-CateninCD68Data adapted fromKeren et al., Cell, 2018VimentinHLA Class ISMAH3K9ac

OPERATES LIKE ATRADITIONAL MICROSCOPEWITH FAST SCAN ANDHIGH RESOLUTION SETTINGSINTUITIVE DATA VISUALIZATIONAND EASY DATA SHARINGInteract withMIBI data atmibi-share.ionpath.com100 µmCOARSEFINEMIBItracker cloud-based datamanagement and visualization platform800x800 µm2800x800 µm21 µm resolution650 nm resolutionMIBItracker software enables review of image quality following arun, assessment of expression profiles across multiple cells, andevaluation of immune cell populations through a web browser.16 min68 minImage files can be exported as TIFFs and readily inserted intopublications or used in subsequent analysis in third-party packagessuch as Fiji, HALO , VisioPharm , and QuPath.

REVOLUTIONARY DATADELIVERED RELIABLYA COMPLETE PLATFORMMULTIPLEXED ION BEAM IMAGING Robust instrumentationtime-of-flight Highly validated reagents Easy-to-use softwaresecondary ionsgeal Im aensionN-Dim40 marker simultaneousdetectionprimary ionsMIBI TECHNOLOGYMIBI Technology is based on secondary ion massspectrometry or SIMS. With SIMS, a primary ion beamis rastered across the surface of a sample, liberatingreporter ions that are then simultaneously recorded ona pixel-by-pixel basis by Time-of-Flight detection. Anion beam, unlike a laser, enables resolution to be tunedover a broad range—in the case of the MIBIScope, from280 nm to 1 micron. Once liberated, the reporter ions,or “secondary ions,” travel uninterrupted at super sonicspeed from the sample to the detector, leading to fastacquisition and extraordinary sensitivity.24/7 DATA GENERATIONPrior to the advent of MIBI technology, SIMSwas primarily used in the semiconductorindustry, where SIMS instruments are reliedon to produce consistent data with roundthe-clock operation. IONpath has leverageddecades of advancements in SIMS todevelop an instrument capable of producingrevolutionary data 24/7.

EASILY INTEGRATES INTOPATHOLOGY WORKFLOWSP R E PA R EetMtalAntigen 2Antigen 3A N A LY Z EAntigen NMIBIMIBIMeMetalAntigen 1talSINGLE STEPIMAGINGIMAGEMeSINGLE STEPSTAININGDETECTal40 MARKERSS TA I NVISUALIZEMIBI is compatible with allcommon sample types, includingFFPE and fresh/frozen tissue.Tissue is stained with metal-taggedantibodies using a standard IHCstaining protocol. All markers arestained in a single step and thesample is stable for months poststaining.The sample is analyzed usingsecondary ion mass spec (SIMS).Low resolution survey scans can becollected prior to high resolutionimaging. The sample is notdestroyed during imaging and itcan be stored for follow-on studiesor utilized for additional analyses.MIBI outputs TIFF images,which can quickly and easilybe viewed on MIBItracker orexported for subsequent analysiswith third-party software.QUANTIFYDISCOVER051015202530

REAGENTSUse our preset panels or label your own antibodiesIO Biomarker PanelsPreset, multiplexed panels permitbroad characterization of the tumormicroenvironment. Each panel has beenextensively validated on a variety oftumor and normal tissue types to ensureoptimal performance across a wide setof sample types.Conjugated AntibodiesSupplement biomarker panels withadditional pre-conjugated antibodies orbuild a unique panel of pre-conjugatedantibodies.Conjugation KitsLabel any antibody of interest for MIBIwith a straightforward protocol that canbe completed in an afternoon.CHECKPOINTPANELEPITHELIAL IOPANELLYMPHOMA CD45CD56CD68CD163dsDNAFOXP3HLA Class 1HLA DRIDO1KeratinKi-67LAG3Na/K zyme BHLA Class 1HLA DRIDO1KeratinKi-67LAG3Na/K IGNdsDNAFOXP3Granzyme BHLA Class 1HLA DRIDO1Ki-67LAG3Na/K ATPasePAX5PD-1PD-L1VimentinContact ath, Inc. 960 O’Brien Drive, Menlo Park, CA 94025For Research Use Only (RUO). Not for diagnostic use. MIBI and MIBIscope are trademarks of IONpath, Inc. in theUnited States or other countries. All rights reserved. 2019 IONpath, Inc. Doc#: MK BR14-001 RevA /11012019

phenotype immune infiltrate MIBIscope A REVOLUTIONARY TECHNOLOGY FOR ANALYSIS OF THE TUMOR MICROENVIRONMENT u mor Cells Immune Cells T cells Helper T cellsCytotoxic T cells Macr ophages B cells Nonpr oliferating T umor Cells 0 10 20 30 40 NUMBER OF CELLS COEXPRESSING PD-L1 COEXPRESSION OF PD