Frozen sections of the aortic root were sectioned using the Leica CM3050S cryostat at 8 µm interval. All images were obtained with a Nikon Microphoto-FXA microscope (Nikon, Tokyo, Japan) equipped with an FX-35-DX- digital camera (Nikon) and ACT-2U imaging software (Nikon). Blinded analysis of the images was performed using image-pro plus software (Media Cybernetics, Rockville (MD), USA).
Act 2u imaging software
Manufactured by Nikon
Sourced in Japan
The ACT-2U imaging software is a tool used for capturing and analyzing microscopic images. It provides a platform for users to acquire, process, and manage digital images obtained from various microscopy techniques. The software's core function is to enable researchers and scientists to document, measure, and evaluate specimens under magnification.
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Lab products found in correlation
Fx 35 dx digital camera, by Nikon (2 mentions)
Microphoto fxa microscope, by Nikon (2 mentions)
Cm3050s cryostat, by Leica (1 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
80i fluorescence microscope, by Nikon (1 mentions)
Hoechst 33258 staining solution, by Beyotime (1 mentions)
Biotinylated secondary antibody, by Vector Laboratories (1 mentions)
3 3 diaminobenzidine tetrahydrochloride dab, by Merck Group (1 mentions)
Horseradish peroxidase conjugated streptavidin, by Merck Group (1 mentions)
Goat serum, by Merck Group (1 mentions)
Eclipse 80i microscope, by Nikon (1 mentions)
Masson s trichrome staining kit, by Merck Group (1 mentions)
5 protocols using act 2u imaging software
1
Aortic Root Histology Imaging
2
Soybean Leaf Vein Development Dynamics
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GmHD-Zip III expression was examined during leaf vein development in soybean, and leaf veins were sampled at 0, 12, 24, 48, and 96 h. Vein tissue development was observed by using cross-sections stained with safranin-O/fast green. The 0.5 cm × 1 cm sections were cut from the central leaflet of six different leaves for each treatment for fixation with FAA. Tissues were embedded in paraffin, and serially, paraffin sections were cut into 10 μm thickness. The sections were stained with safranin-O/fast green. Sections were scanned under a Nikon Eclipse 80i microscope, and images were acquired with the ACT-2U imaging software (Nikon Corporation) [73 (link)].
3
Apoptosis Analysis of H9c2 Cells
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H9c2 cells were seeded on sterile cover glasses placed in 24-well plates and cultured in Dulbecco’s Modified Eagle medium. Upon reaching 50–60% confluence, cells were treated with the above drugs (ISO, ISO + PUE, and ISO + PUE@PEG-PE). After treatment for 24 h, cells were fixed, washed twice with PBS, and stained with Hoechst 33258 staining solution according to the manufacturer’s instructions (Beyotime, Haimen, Jiangsu, China). Image capture and slide evaluations were performed using a Nikon 80i fluorescence microscope equipped with ACT-2 U imaging Software (Nikon, Tokyo, Japan). Apoptotic cells were defined by the condensation of nuclear chromatin, fragmentation, or margination to the nuclear membrane.
4
Immunohistochemical Analysis of Tau Pathology
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Fixed (4% paraformaldehyde) free-floating coronal brain sections (40 μm thickness) from 9 month-old ThyTau22 and P301S mice were assayed. After antigen retrieval (80°C for 20 min in 50 mM citrate buffer, pH 6.0), endogenous peroxidase was inhibited (3% H2O2/10% methanol in PBS, pH 7.4 for 20 min) and non-specific staining was avoided using 5% goat serum (Sigma-Aldrich) in PBS. Sections were incubated with the primary antibody (overnight at room temperature) followed by the corresponding biotinylated secondary antibody (1:500 dilution, 1 h at room temperature, Vector Laboratories), streptavidin-conjugated horseradish peroxidase (1:2000, 90 min, Sigma-Aldrich), and visualized with 0.05% 3-3-diaminobenzidine tetrahydrochloride (DAB, Sigma-Aldrich) and 0.01% hydrogen peroxide in PBS. The specificity of the immune reactions was controlled by omitting the primary antisera. Sections were examined under a Nikon Eclipse 80i microscope and images were acquired with a Nikon DS-5M digital camera using the ACT-2U imaging software (Nikon Corporation).
5
Histological Analysis of Aortic Lesions
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Frozen sections (8 μm) of aortic sinus lesions were prepared, and stained for neutral lipid accumulation in lesion sections by Oil red O staining. Collagen content was measured by staining with Masson's trichrome staining kit (HT15‐1KT; Sigma–Aldrich). Necrotic core area was quantified by assessing unstained area in plaques stained with Masson's trichrome. Nuclei for all immunohistochemistry stains were counterstained using Gill's Hematoxylin. Immunohistochemistry stains were visualized with a Nikon Microphoto‐Fxa microscope (Nikon) equipped with an FX‐35‐DX digital camera (Nikon). Images were captured using 4x/0.13 objective lens using the ACT‐2U imaging software (Nikon). Image quantification was performed using Image Pro Plus software. For en face analysis, descending aortas were fixed in 10% formalin, trimmed of fat, stained with Oil red O, opened longitudinally, pinned out on black wax, and digitally photographed, as described.11 Images were captured using a dissecting photomicroscope. Image quantification for en face analysis was performed using Biopix software (Gothenburg, Sweden).
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