Figure 6.
Figure 6. Laser confocal microscopy characterization of PVR- and Nectin-2-expressing cells in human lymph nodes. Human lymph nodes were stained for mannose receptor (MR; red) and PVR (A) or Nectin-2 (D; green) and were analyzed by confocal microscopy. Cells around the HEV show colocalization of DNAM-1 ligands and MR (merged images, yellow), whereas single staining for DNAM-1 ligands is visible on endothelial cells. High-power magnifications focused on single cells around the HEVs show costaining of MR and CD11c markers with PVR (B-C) and Nectin-2 (E-F). No staining was observed by using an isotype-matched control mAb with irrelevant specificity (not shown). Bar indicates 20 μm. Images in panels A-E were acquired using an LSM 510 Meta confocal microscope (Zeiss, Jena, Germany) equipped with a 40 ×/1.30 NA oil Plan-Neofluor objective lens (Zeiss). Images in panel F were acquired using an LSM 510 Meta laser scanning confocal microscope equipped with a 63 ×/1.4 NA oil Plan-Neofluor objective lens (Zeiss). LSM 510 Meta confocal microscope software version 3.0 (Zeiss) was used to capture all the images.

Laser confocal microscopy characterization of PVR- and Nectin-2-expressing cells in human lymph nodes. Human lymph nodes were stained for mannose receptor (MR; red) and PVR (A) or Nectin-2 (D; green) and were analyzed by confocal microscopy. Cells around the HEV show colocalization of DNAM-1 ligands and MR (merged images, yellow), whereas single staining for DNAM-1 ligands is visible on endothelial cells. High-power magnifications focused on single cells around the HEVs show costaining of MR and CD11c markers with PVR (B-C) and Nectin-2 (E-F). No staining was observed by using an isotype-matched control mAb with irrelevant specificity (not shown). Bar indicates 20 μm. Images in panels A-E were acquired using an LSM 510 Meta confocal microscope (Zeiss, Jena, Germany) equipped with a 40 ×/1.30 NA oil Plan-Neofluor objective lens (Zeiss). Images in panel F were acquired using an LSM 510 Meta laser scanning confocal microscope equipped with a 63 ×/1.4 NA oil Plan-Neofluor objective lens (Zeiss). LSM 510 Meta confocal microscope software version 3.0 (Zeiss) was used to capture all the images.

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