The eyeball and dissected lymph nodes were perfused with PBS followed by 4% paraformaldehyde overnight to be processed for paraffin embedding. Five-micrometer sections were cut from the paraffin blocks, mounted on glass slides, and labeled with mouse anti- rat CD3 (DB-082; DB Biotech) and anti-cytomegalovirus antigen (BSB 5454; Bio SB). A Horseradish Peroxidase Labeling Kit (TAHC03D; BioTnA, Kaohsiung City, Taiwan) was used for detect diaminobenzidine staining. Paraffin sections were stained hematoxylin and eosin staining. Images were obtained with MoticEasyScan Pro (Motic Asia, Hong Kong) and processed with EasyScanner software.
Moticeasyscan pro
Manufactured by Motic
Sourced in Hong Kong, Canada, China
The MoticEasyScan Pro is a compact, high-resolution digital slide scanner designed for laboratory applications. It captures high-quality digital images of microscope slides, enabling efficient digitization and archiving of specimens.
Automatically generated - may contain errors
4 protocols using moticeasyscan pro
1
Immunohistochemical Analysis of Lymphoid Tissues
2
Immunohistochemical Analysis of Mouse Brain
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The brain tissues were collected from suckling mice and fixed with 10% formaldehyde for 48 h. The tissues were embedded in paraffin and cut into 3 μm-thick sections on slides for Hematoxylin/Eosin (H&E) and immunohistochemical (IHC) staining. The tissues were incubated with rabbit anti-ENO1 (1:100; GeneTex, CA, USA) or rabbit anti-EV-A71 (1:500; GeneTex, CA, USA) antibodies at 37 °C for 1 h and then with Goat anti-Rabbit IgG (H + L) as a secondary antibody (1:50; Leadgene Biomedical, Tainan, Taiwan) at room temperature for 30 min. The stained slides were photographed using a light microscope equipped with a Motic EasyScan Pro (NA 0.75, Canada) for morphometric analysis.
3
Lung Histomorphometry Analysis Protocol
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Lung samples were inflated with 10% neutral-buffered formalin by intratracheal instillation at a pressure of 25 cmH2O for 10 min. The lung tissues were then embedded in paraffin and cut into sections for staining with haematoxylin and eosin (H&E). Lung H&E images were obtained by Motic Easyscan Pro and Motic DSAssistant software (Motic, Xiamen, Fujian, China). The thickness of the airway wall was assessed through the use of ImageJ software (National Institute of Health, Bethesda, MD, USA) based on images captured at a 20x magnification. The MLI assessed the length of lines crossed and the number of grid lines intercepts in alveolar space from 10 non-overlapping fields of each lung sample according to previously described method (Crowley et al., 2019).
4
Automated Immunohistochemistry Staining Protocol
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H&E-stained slides of the most invasive part of primary tumor were used to determine the DR types. All these slides were scanned using digital WSI scanning systems (Aperio AT2, Leica; Aperio GT 450 Leica; MoticEasyScan Pro, Motic; KF-PRO-020, KFBIO; SQS-600P, TEKSQRAY; NanoZoomer S60) at 40 × magnification (resolution: 0.21–0.26 μm /pixel). Image annotation was carried out using the ImageScope software (ImageScope v12.4.3, Leica). Subsequently, 620 slides were selected for IHC. A series of steps were performed, deparaffinage, antigen retrieval solution (using 10 × concentrate solution, Novocastra, Leica) and primary [human anti-CD3 (Gene Tech, catalog no. GT200229) rabbit mAbs] and secondary (rabbit-anti-mouse IgG, Bond Refine Detection Kit, Leica) antibodies, according to the manufacturer's recommendations in a Ventana BenchMark automated staining system. Finally, the sections were incubated with 3,3-Diaminobenzidine, counterstained with hematoxylin, and mounted using special glue. To guarantee quality assurance, an internal positive control was utilized. The IHC-stained tissue sections were then captured utilizing a digital whole-slide scanning system (Aperio AT2, Leica) at 40 × magnification.
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