Tissue specimens for each patient were cut and stained using anti-CD3 (clone SP7, Ventana Medical Systems, Roche), anti-CD20 (clone L26, Ventana Medical Systems, Roche) and anti-CD30 (clone Ber-H2,Ventana Medical Systems, Roche) in the Pathology departments of the participating hospitals. Subsequently, scanning of the slides was centralized at the Puerta del Mar University Hospital (Cadiz, Spain). The PANNORAMIC ® 250 Flash III DX scanner (3D Histech Ltd., Budapest, Hungary) was used to perform the scanning of tissue slides with a resolution of 0.25 microns per pixel, obtaining digital slides in MRXS file format, according to the manufacturer’s recommendations. SlideViewer 2.5 software (3D Histech Ltd., Budapest, Hungary) was used to review the digital slides.
The 3DHistech QuantCenter application (version 2.3) was used for the quantification of CD3, CD20 and CD30 positive cells. Cell quantification was performed in the whole lymph-node specimen, excluding unstained areas and artifacts. Consequently, cell quantification was not restricted to areas of interest within the specimen, and data obtained reflect the whole composition of the diagnostic samples. See theSupplementary Material
The 3DHistech QuantCenter application (version 2.3) was used for the quantification of CD3, CD20 and CD30 positive cells. Cell quantification was performed in the whole lymph-node specimen, excluding unstained areas and artifacts. Consequently, cell quantification was not restricted to areas of interest within the specimen, and data obtained reflect the whole composition of the diagnostic samples. See the