Fig. 2: Tissue microarray samples collected from the tumor center and invasive front were immunostained to determine microsatellite status, the consensus-molecular-subtype (CMS1 (MSI), CMS2/3 and CMS4), and the immunoscore. | Modern Pathology

Fig. 2: Tissue microarray samples collected from the tumor center and invasive front were immunostained to determine microsatellite status, the consensus-molecular-subtype (CMS1 (MSI), CMS2/3 and CMS4), and the immunoscore.

From: Associations of non-pedunculated T1 colorectal adenocarcinoma outcome with consensus molecular subtypes, immunoscore, and microsatellite status: a multicenter case-cohort study

Fig. 2

a For each tumor specimen, H&E stained slides containing tumor center (TC) and invasive margin (IM) regions were selected by two expert pathologists. b For each region, three cores (diameter, 0.6mm) were punched out and were set into a recipient paraffin block using an automated tissue microarrayer (TMA Grand Master 3D Histech, Sysmex). c TMA core stained with antibodies against CD3 (tumor in blue; CD3 in brown). d The density (number of positive cells/surface area analyzed) of lymphocytes within the tumor center and invasive margin was quantified with QuPath image analysis. e The immunoscore was based on the enumeration of two lymphocyte populations (CD3 and CD8). The consensus-molecular-subtype was based on a panel of five IHC stains (CDX2, FRMD6, HTR2B, ZEB1, KER) together with microsatellite status. #Formula for immunoscore in non-pedunculated T1 CRCs: (((log(CD3 IM+6.35)−6.5185)/0.9117)+((log(CD3 TC+19.325)−6.4082)/0.8030) + ((log(CD8 IM+0.3333)−4.4885)/1.4498) + ((log(CD8 TC+0.5)−4.1358)/1.4636))/4. If the result is≤-0.64175 patient is I-Low. If the result is >-0.64175 patient is I-High. CD3 IM = CD3 density at invasive margin; CD3 TC = CD3 density at tumor center; CD8 IM = CD8 density at invasive margin; CD8 TC = CD8 density at tumor center.

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