Immunostaining of knee joint sections was performed with specific antibodies against target proteins according to a previously published protocol with some modifications (Wang et al. 2019 (link)). Sections were incubated with anti-NLRP3 (1:100), anti-ASC (1:100), anti-caspase-1 (Wanleibio, 1:100), anti-GSDMD (Affinity Biosciences, 1:100), and anti-IL-1β (1:100) antibodies overnight at 4 °C. On the following day, these tissue sections were incubated with a polymer-HRP detection system (PV9001, ZSGB-BIO) and visualized with a diaminobenzidine (DAB) peroxidase substrate kit (ZLI-9017, ZSGB-BIO). Each section was evaluated under a microscope (DMi8, LEICA) at 20 × magnification. Histological analyses were performed in a blinded manner by two independent observers. Image-ProPlus6 software 6.0 (Media Cybernetics, Inc.) was used to analyze the average integrated optical density (IOD) of three randomly selected areas of the acquired images.
Image proplus6 software 6
Manufactured by Media Cybernetics
Image-ProPlus6 software 6.0 is an image analysis and processing software developed by Media Cybernetics. The software provides tools for capturing, processing, analyzing, and managing digital images.
Automatically generated - may contain errors
2 protocols using image proplus6 software 6
1
Immunostaining Analysis of Knee Joint Proteins
2
Immunostaining of Knee Joint Proteins
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Immunostaining of knee joint sections was performed with speci c antibodies against target proteins according to a previously published protocol with some modi cations [29] . Sections were incubated with anti-NLRP3 (1:100), anti-ASC (1:100), anti-caspase-1 (Wanleibio, Wuhan, China, Cat. # WL02996, 1:100), anti-GSDMD (A nity Biosciences, Beijing, China, Cat. # AF4012, 1:100), and anti-IL-1β (1:100) antibodies overnight at 4°C. On the following day, these tissue sections were incubated with a polymer-HRP detection system (PV9001, ZSGB-BIO, Beijing, China) and visualized with a diaminobenzidine (DAB) peroxidase substrate kit (ZLI-9017, ZSGB-BIO, Beijing, China). Each section was evaluated under a microscope (DMi8, LEICA, Wetzlar, Germany) at 20 X magni cation. Image-ProPlus6 software 6.0 (Media Cybernetics, Inc.) was used to analyze the average integrated optical density (IOD) of three randomly selected areas of the acquired images.
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