Mouse brain samples were perfused with Tyrode solution followed by PFA, then post‐fixed in 4% PFA for 48 h at 4 °C, and paraffin‐embedded at the University of Michigan's facility using Leica ASP 300 and Tissue‐Tek.[26 (link),
28 (link)
] Sections (5 µm) were obtained using a Leica rotary microtome. For histopathological examination, sections underwent Hematoxylin and Eosin (H&E) staining. Concurrently, DAPI staining was performed for Immunofluorescence (IF) in GFP‐positive cells. Imaging of H&E‐stained sections utilized an Olympus BX53 Upright Microscope, selecting ten random fields per section to encompass tumor heterogeneity. For immunofluorescence, paraffin‐fixed glass coverslips post oncostreams formation was imaged using a Zeiss LSM 880 laser scanning confocal microscope. The resulting data were analyzed using the Zeiss Zen (Blue edition) software, version 2.5.
28 (link)
] Sections (5 µm) were obtained using a Leica rotary microtome. For histopathological examination, sections underwent Hematoxylin and Eosin (H&E) staining. Concurrently, DAPI staining was performed for Immunofluorescence (IF) in GFP‐positive cells. Imaging of H&E‐stained sections utilized an Olympus BX53 Upright Microscope, selecting ten random fields per section to encompass tumor heterogeneity. For immunofluorescence, paraffin‐fixed glass coverslips post oncostreams formation was imaged using a Zeiss LSM 880 laser scanning confocal microscope. The resulting data were analyzed using the Zeiss Zen (Blue edition) software, version 2.5.
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