Slides containing sections obtained at 100-µm intervals (i.e., every 10th section) were fixed in Bouin’s Solution at 55°C for 1 h and then treated with Weigerts iron hematoxylin working solution for 10 min, with Biebrich scarlet-acid fuchsin for 5 min, with phosphomolybdic acid-phosphotungstic acid for 5 min, with aniline blue solution for 5 min, and with 1% acetic acid for 1 min; then, the sections were dehydrated in 95% alcohol for 2 min, cleared with 2 changes of xylene, and mounted with permount and coverslips overnight. Sections were imaged with a ×10 objective on an Olympus B×51 Fluorescence Microscope; when multiple fields of view were required for a single section, the images were stitched together with Photoshop software.
B 51 fluorescence microscope
Manufactured by Olympus
The B×51 Fluorescence Microscope is an optical microscope designed for fluorescence imaging applications. It features a high-intensity illumination system and advanced optics to enable visualization and analysis of fluorescently labeled samples.
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Lab products found in correlation
3 protocols using b 51 fluorescence microscope
1
Collagen Fiber Staining Protocol
2
Cardiomyocyte Apoptosis Analysis
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Apoptotic cardiomyocytes were detected in 7-μm-thick myocardial cryosections using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining following the manufacturer’s protocol (Thermo Fisher Scientific, New Zealand). Following this, sections were probed with cardiac troponin mouse monoclonal antibody (1:100 dilution, Novus Biologicals, USA) to label cardiomyocytes and counterstained with Alexa Fluro 633 goat-anti-mouse secondary antibody (1:1,000 dilution, Thermo Fisher Scientific, New Zealand). Images were captured at 200× magnification using an Olympus B×51 fluorescence microscope. Data are expressed as the percentage of TUNEL-positive cardiomyocytes in the section.
3
Adhesion Assay for Cancer Cells
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Cancer cell-endothelial adhesion was performed as previously described (10 (link)). A549 cells were cultured in 6-well plates with different concentrations (0.0, 2.5, 5.0 and 10.0 µM) of 7a and HUVECs were cultured in 24-well plates. Following incubation at 37°C for 48 h, A549 cells were washed with PBS and labelled using 5 µM DiO fluorescent cell labelling solution in serum-free RPMI-1640 medium for 30 min at 37°C. Next, tumor cells were washed with PBS and removed from the culture plates. A549 cells were collected using centrifugation at 150 × g for 3 min at room temperature. Following washing, 5×104 cells were applied to the HUVEC monolayer cultured in 24-well plates for 1 h at 37°C. Next, the 24-well plates were gently washed with PBS and the fluorescently labelled cells were counted in 10 randomly selected fields using an Olympus B51 fluorescence microscope (Olympus Corporation) with a 10 objective (magnification, ×100).
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