To analyze HBcAg expression digital images were acquired with an AxioImager.MI microscope (Zeiss, 20 Germany) using an in-house Metamorph macro (Molecular Devices, USA), that allows capture of randomly located images at 20X (Plan-Neofluar objective with 0.50 NA), with automatic focus, white balance and shadow correction, on every acquired image. All images were stored in uncompressed 24-bit color TIFF format. Images were automatically analyzed using a plugin developed by Fiji, ImageJ [24 ].
Metamorph macro
Manufactured by Molecular Devices
Sourced in United States
MetaMorph Macro is a powerful software tool designed to automate image acquisition and analysis workflows. It provides a comprehensive set of features to streamline various imaging tasks, including image capture, processing, and analysis. The core function of MetaMorph Macro is to allow users to create customized, automated scripts that can be applied to their imaging experiments, enabling consistent and efficient data collection and analysis.
Automatically generated - may contain errors
3 protocols using metamorph macro
1
Automated HBcAg Expression Imaging
2
Automated F4/80 Expression Imaging
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To analyse F4/80 expression digital images were acquired with an AxioImager. MI microscope (Zeiss, Germany) using an in-house Metamorph macro (Molecular Devices, USA). All images were stored in uncompressed 24-bit colour TIFF format. Images were automatically analysed using a plugin developed for Fiji, ImageJ.41.
3
Histological and Molecular Analysis of Liver Tissue
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Liver sections were fixed in 4% paraformaldehyde (Panreac), embedded in paraffin, sectioned (thickness, 5 μm), and stained with hematoxylin and eosin (H&E). Immunohistochemistry (IHC) for HBcAg (B0586; Dako, Glostrup, Denmark) and PD-L1 (ab58810; Abcam) was performed using the EnVision system (Dako) according to the manufacturer's recommendations. In situ detection of cells with DNA double-strand breaks was performed by the terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining method by use of an In Situ Cell Death Detection kit (Roche) according to instructions. Images were acquired digitally with an Axio Imager. MI microscope (Zeiss, Germany) using an in-house MetaMorph macro (Molecular Devices, USA). All images were stored in uncompressed 24-bit color TIFF format. Images were automatically analyzed using a plugin developed for Fiji, ImageJ, at NIH.
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