At sacrifice, different adipose tissue depots, including visceral white adipose tissue (vWAT), inguinal WAT (iWAT), and interscapular brown adipose tissue (BAT), were collected and weighed. Then, vWAT, iWAT, and BAT were fixed with 4% (vol/vol) paraformaldehyde/PBS for 24 h at 4 °C, embedded in paraffin, sectioned, and stained with hematoxylin/eosin (H&E). The Mean adipocyte area was evaluated in sections of vWAT, iWAT, and BAT using Image J software (version 1.52, realized by National Institutes of Health, USA). Moreover, iWAT sections were processed for staining for UCP-1 (1:2000, Abcam, Cambridge, MA, USA). The reaction was revealed with the Dako EnVisionTM FLEX+ detection system (Dako Italia S.p.A., Milan, Italy). Image scanning of the histological samples was performed using Nanozoomer and acquired using NDP.view2 Image viewing software (Hamamatsu, Shizuoka, Japan). The brown area was measured using a photoshop instrument (Photoshop, version 25.0, by Adobe, San Jose, CA, USA) that led to the selection of the brown area in figures with the same magnification and area.
Ndp view2 image viewing software
Manufactured by Hamamatsu Photonics
Sourced in Japan
NDP.view2 is an image viewing software developed by Hamamatsu Photonics. It is designed to display and analyze digital images captured with Hamamatsu's scientific cameras and imaging devices.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using ndp view2 image viewing software
1
Adipose Tissue Histomorphometry and UCP-1 Analysis
2
Slide Analysis using Light Microscopy
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Slides were mounted and analyzed using light microscopy. Microphotographs were taken after slide digitalization using the Nano Zoomer S210 (C13239-01, Hamamatsu Photonics, Hamamatsu, Japan) and the NDP.view2 image viewing software (U12388-01, Hamamatsu Photonics, Hamamatsu, Japan). Evaluation was performed independently by two investigators. Statistical methods were not applied since the study was descriptive.
3
Histopathological Evaluation of Suspicious Lesions
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Patients with suspicious VLS based on clinical manifestations and well‐established dermoscopic criteria5 received a skin biopsy at the same lesion site where HFUS examination was performed. The histopathological features, including epidermal atrophy, follicular plug, homogenization of collagen in the superficial dermis, and lichenoid lymphocytic infiltration, were further recorded. The maximum thickness of collagen homogenization and lymphocytic infiltration were also measured using the length measurement tool of NDP.view2 image viewing software (HAMAMATSU PHOTONICS K.K., Hamamatsu City, Japan). The histopathology was examined by a dermatopathologist with more than 5‐year experience.
4
Histological Analysis of MucilAir Nasal Cultures
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MucilAir ™ nasal cultures were fixed at 4% paraformaldehyde in pH 7.4 phosphate buffered saline for 20 h and transferred to 70% ethanol/water and shipped to Cerba Research (Montpellier, France). For further processing, tissue samples were incubated with 10% neutral buffered formalin (NBF), various alcohol dilutions, xylene, and paraffin for a total of one and a half hours. After processing samples, tissues were embedded in paraffin for a permanent state of preservation. Tissues were sectioned at 4 µm, with 1 section per slide. After sectioning, samples were placed in an oven for additional adhesion. Slides were stained with hematoxylin and eosin (H&E) using reagents from Leica and Statlab ™ . After staining, sections were dehydrated, and the film cover slipped using a Tissue-Tek Film ™ automated Coverslipper. The whole slide scanning (×40 objective) was performed on an Aperio AT2 digital whole slide scanner (Leica Biosystems). Scanned images of slides for the MucilAir ™ nasal mocktreatment (air-only) VITROCELL ® chamber controls were compared to incubator controls from 5-day and 15-day studies. Images were viewed using Hamamatsu NDP.view2 image viewing software and evaluated by a board-certified veterinary pathologist (S Elmore).
5
Immunostaining and Scoring of FHOD1, INF2, and DAAM1
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TMAs were sectioned at 3.5 μm and stained with rabbit anti-human FHOD1 (1:500; Sigma-Aldrich, USA), INF2 (1:500; Proteintech, USA), and DAAM1 (1:200; Proteintech). Immunostaining was performed according to the streptavidin-peroxidase method, using a Labvision staining device (Thermo Fisher Scientific, USA). Stained slides were scanned using a NanoZoomer S60 slide scanner (Hamamatsu Photonics, Japan), and a pathologist (MG) and researcher (MP) evaluated the staining intensity using the NDP.view2 Image viewing software (Hamamatsu).
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