Liver fibrosis was evaluated using Van Gieson staining. Briefly, paraffin-embedded liver sections were deparaffinized and hydrated to distilled water first and stained with Wright’s Working Hematoxylin for 10 min and then washed in distilled water. The slides were then stained with Van Gieson solution for 3 min, followed by gradient dehydrating in 95% alcohol, absolute alcohol, and 2 changes in xylene before mounting with DPX. These slides were scanned through slide scanner, Zeiss Axio ScanZ1 (Carl Zeiss Ltd. Cambridge, UK), and the whole field image of each section was processed via ZEN 2010 blue edition software (Carl Zeiss Ltd. Cambridge, UK) and quantified for positive staining for the fibrotic area (pink) using ImageJ analysis software (NIH, USA) and an automated macro. The proportionate area of fibrosis staining (pink) was calculated using: Pixels of area that stained pink / Pixels of the whole image.
Zen 2010 blue edition software
Manufactured by Zeiss
Sourced in United Kingdom, United States
The ZEN 2010 blue edition software is a comprehensive digital imaging and processing platform designed for microscopy applications. It provides a user-friendly interface for capturing, analyzing, and managing microscopic images. The software offers core functionalities for image acquisition, processing, and data management, enabling researchers and scientists to effectively utilize their microscopy equipment.
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2 protocols using zen 2010 blue edition software
1
Liver Fibrosis Quantification via Van Gieson Staining
2
Neuronal Morphology Analysis Protocol
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Neurons were fixed for 10 min with 4% paraformaldehyde in phosphate-buffered saline (ThermoFisher Scientific, #J61899-AP, USA) and stained as previously described (32 (link)). In brief, neurons were immunolabeled using primary antibodies against MAP2ab (Millipore, MAB378, USA; 1:500) with secondary antibodies conjugated to goat-anti-mouse Alexa 488 (ThermoFisher Scientific, #O-6380, USA; 1:1,000) diluted in PBS (ThermoFisher Scientific, #20012043). Nuclei of cells were stained using Hoechst 33342 (3 min; ThermoFisher Scientific, #H3570, USA) and coverslips were mounted using Prolong Gold (ThermoFisher Scientific, #P36930, USA). Z-stack images were obtained using ZEN 2010 Blue Edition software (Zeiss, Thornwood, NY, USA) with a Zeiss LSM 700 laser scanning confocal microscope using a 63x immersion objective (Zeiss, Thornwood, NY, USA). Dendritic branching complexity (e.g., maximum process length and distance from soma with maximal branching) and soma area were assessed with orthogonal projections from Z-stack images using the Sholl analysis tool within ImageJ software [Version 2.1.0; (63 (link))].
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