Ndp view2 viewing software u12388 01

Three points (dorsal, lateral, and ventral) from each brain and hemisphere were selected for cortical thickness^{35 (link)}. The most rostral section measured was located at ∼2.10 mm anterior to bregma and the most caudal section at ∼−0.46 mm posterior to bregma. For each point, a vector was considered from the tangent of the outer edge to the inner edge of the cortex. The NDP.view2 viewing software U-12388-01 (Hamamatsu, Japan) was used to record up to six measurements of cortical thickness from each coronal section, three from each hemisphere. Cortical thickness in the present experiment was a mean measure of both hemispheres in seven consecutive slices.

Approximately 3-4 mm² pieces of tissue were collected at the same sites sampled for gene expression analysis and tissues were immediately embedded in OCT (Optimal cutting temperature compound) and stored in sealed containers at -80°C. Tissue sections (8 μm) were cut for hematoxylin and eosin (H&E) staining, acid fast staining, and immunohistochemical staining at the Plateforme d’histologie et microscopie électronique, Faculté de médecine, Université de Sherbrooke.
For immunohistochemistry, the primary antibodies were polyclonal rabbit anti-human CD3 (clone A0452, 1:400 dilution) (DAKO, Glostrup, Denmark), anti-bovine CD11c mAb (clone BAQ153A, 1:50 dilution)] (Kingfisher Biotech, Saint Paul, MN, USA), mouse anti-bovine CD172a mAb (clone CC149, 1:500 dilution) (Bio-Rad Laboratories, Mississauga, ON, Canada), mouse anti-Ki67 (clone MIB-1, 1:50 dilution) (Agilent Technologies, Mississauga, ON, Canada), and a purified mouse IgG1 isotype control (product code MG100, 1:50 dilution) (Life Technologies, by Thermo Fisher Scientific, Mississauga, ON, Canada). Histological images of the stained tissues were acquired using a slide scanner (NanoZoomer Digital Pathology, Hamamatsu Photonics, Boston, MA, USA) followed by viewing with NDP.view2 viewing software U12388-01 (Hamamatsu Photonics, Boston, MA, USA).

HP, aHP, and PBS were intraperitoneally injected into Wistar rats after depilation of the back skin at a porphyrin concentration of 2 mg kg⁻¹. Twenty-four hours after drug injection, the back skin was irradiated with ultraviolet A (UVA) at 385 nm at 10 J cm⁻². Twenty-four hours after UVA irradiation, the skin on the back was collected under anesthesia with isoflurane. The tissues were then fixed in 10% formalin buffer solution at pH 7.4, and tissue cross-sections were histologically observed using hematoxylin and eosin (HE) staining. The images were scanned using a digital slide scanner (NanoZoomer S210, Hamamatsu Photonics, Hamamatsu, Japan), and the hydrogel layer area was quantified using the NDP.view2 Viewing software U12388-01 (Hamamatsu Photonics, Hamamatsu, Japan).