Masson trichrome-stained collagen fibers were visualized by light microscopy (Zeiss, Cyber-Shot, Japan). To assess the collagen intensity in the histological sections, the pixel-based intensity of blue-staining, representing the collagen fibers, was assessed at 2530 µm × 2530 µm sections of a photomicrograph by software analysis (Image pro-insight, Media Cybernetics, USA). For this purpose, 20-megapixels images were prepared by the on-board camera (Zeiss, Cyber-Shot, Japan), then, the area (2530 µm × 2530 µm) was measured by MEZZURE software on the photomicrograph, and the means of pixel-based intensities, obtained from 3 images from a section (in total 15 section/each group), were evaluated. Finally, the mean ± SD of intensities was compared between groups34 .
Cyber shot
Manufactured by Zeiss
Sourced in Japan
The Cyber-Shot is a high-performance laboratory microscope designed for precise and detailed imaging. It features advanced optics and camera technology, providing users with clear and accurate visual data for their research and analysis needs.
Automatically generated - may contain errors
5 protocols using cyber shot
1
Quantitative Analysis of Collagen Fibers
2
Histopathological Analysis of Testis
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After excision of the left testis, it was placed in 10% formalin for 48 hr to fix its tissue. After tissue processing, 5 µm thick sections were prepared with a rotary microtome (Leica Model RM 2145, Germany). Then the slides were stained with hematoxylin and eosin (H&E) (Merck, Germany) and observed under a light microscope equipped with a Sony camera (Zeiss, Cyber-Shot, Japan) (21). The number of spermatogonia, primary spermatocytes, spermatid, and Leydig cells was examined in the histopathological analysis.
3
Assessment of Toothpaste Cytotoxicity on Gingival Fibroblasts
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Human gingival fibroblasts (hGF) were routinely cultivated in DMEM supplemented with 10% fetal bovine serum, penicillin, and streptomycin at 37 o C and 5% CO 2 . The cells were grown at a density of 2x10 4 in 24-well plates and incubated for 24 hours at 37 o C and 5% CO 2 . The cell cultures were then exposed to different dilutions (1:1, 1:2, 1:4, and 1:8) of the toothpaste solutions for 24 hours and fixed in 4% formaldehyde thereafter. Then, Fluoroshield with DAPI (Sigma-Aldrich) and phosphate-buffered saline (PBS, Cultilab, Campinas, SP, Brazil) were added to the wells, which were photographed with a digital camera (Sony F828 Digital, CyberShot, 8.0 megapixels) coupled to an inverted light microscope (Carl Zeiss Microscope Micro lmaging GmbH -Axiovert 40C, Germany). At least 10 photos in different fields were taken in each well. A cell counter (Image J software) was used to help with the count of micronuclei. The number of micronuclei was determined microscopically in 2,000 cells/well and the differences between the median values were statistically analyzed using the Mann-Whitney U test and two-way-ANOVA at a 5% significance level.
4
Collagen Visualization via Masson's Trichrome
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Using Masson’s trichrome stain, collagen fibers were
visualized by light microscopy (Zeiss, Cyber-Shot, Japan)
using the MEZZURE software (Image pro-vision insight
software) with a ×2.4 optical zoom. Staining intensity and
distribution were evaluated by pixel counting.
visualized by light microscopy (Zeiss, Cyber-Shot, Japan)
using the MEZZURE software (Image pro-vision insight
software) with a ×2.4 optical zoom. Staining intensity and
distribution were evaluated by pixel counting.
5
Quantifying Uterine NK Cells in RIF
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Immunohistochemistry (IHC) with pre-diluted anti-CD56 (clone 123C3) mouse monoclonal primary antibody (Ventana Medical Systems, Roche Diagnostics, Germany) was performed on twenty endometrial biopsies obtained from ten RIF patients to detect uterine NK cells (uNKs). CD56-positive cells were counted per one mm
2of the tissue slide. Images were taken and prepared with an on-board camera (Cyber-Shot, Zeiss, Japan). Scale bars were produced using a morphometric lens (CH-2, Olympus, Germany) and proofed with Meazure software (C Thing Software, Sunnyvale, CA, USA). All images were captured under 100 × magnification. Pixel-based frequency values were obtained using Image-Pro-insight software (Version 9.0, Media Cybernetics, USA).
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Immunohistochemistry (IHC) with pre-diluted anti-CD56 (clone 123C3) mouse monoclonal primary antibody (Ventana Medical Systems, Roche Diagnostics, Germany) was performed on twenty endometrial biopsies obtained from ten RIF patients to detect uterine NK cells (uNKs). CD56-positive cells were counted per one mm
2of the tissue slide. Images were taken and prepared with an on-board camera (Cyber-Shot, Zeiss, Japan). Scale bars were produced using a morphometric lens (CH-2, Olympus, Germany) and proofed with Meazure software (C Thing Software, Sunnyvale, CA, USA). All images were captured under 100 × magnification. Pixel-based frequency values were obtained using Image-Pro-insight software (Version 9.0, Media Cybernetics, USA).
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