Imaging analyses were performed using the Opal multiplex tissue staining system and the Mantra quantitative pathology workstation (PerkinElmer, Waltham, MA). Slides from biopsied tumor tissues were stained using an Opal multiplex tissue staining system (PerkinElmer, Waltham, MA). Antigen retrieval was performed by heating slides to 93 ± 2°C for 20 minutes in a high-pH antigen unmasking solution (H-3301, Vector Labs, Burlingame, CA), followed by blocking in 5% bovine serum albumin (BSA) (Jackson ImmunoResearch, Birmingham, AL) in phosphate-buffered saline (PBS). Cell phenotyping and counting were performed using the Mantra quantitative pathology workstation (PerkinElmer) within representative fields preselected by trained hematopathologists. Spatial distribution and marker intensity in target cells were analyzed using inForm® image analysis (PerkinElmer) and Spotfire (TIBCO Software, Palo Alto, CA) software.
Miyawaki K., Kato K., Sugio T., Sasaki K., Miyoshi H., Semba Y., Kikushige Y., Mori Y., Kunisaki Y., Iwasaki H., Miyamoto T., Kuo F.C., Aster J.C., Ohshima K., Maeda T, & Akashi K. (2022). A germinal center–associated microenvironmental signature reflects malignant phenotype and outcome of DLBCL. Blood Advances, 6(7), 2388-2402.
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