Snap-frozen LV tissue was fixed in 4% PFA and embedded in paraffin. Mid-ventricular sections of 7 µm thickness were taken on Superfrost Plus glass slides (Fisher) using a Leica RM2255 microtome. These slides were then stained with Picrosirius Red, Fast Green, and Alcian Blue (RGB) trichrome stain as preciously described40 (link), to visualize collagen protein in the sections. Images were obtained using an Olympus BX51 upright microscope with a Leica DFC7000T camera and Leica Application Suite v4.6 imaging software. Areas with collagen, based on colour thresholds, were detected automatically from RGB trichrome images by a custom Fiji ImageJ 1.53c (NIH, USA) macro, which calculated percent fibrosis. The same macro was applied to all images from both genotypes at both timepoints.
Bx51 upright microscope
Manufactured by Leica
The BX51 upright microscope is a high-performance optical instrument designed for advanced imaging and analysis. It features a sturdy, ergonomic construction and delivers exceptional optical performance, making it suitable for a wide range of applications in research and clinical settings.
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Lab products found in correlation
M165fc stereoscope, by Leica (2 mentions)
Dfc7000 t camera, by Leica (1 mentions)
Superfrost plus glass slide, by Thermo Fisher Scientific (1 mentions)
Tcs sp5, by Leica (1 mentions)
Dm6000b microscope, by Leica (1 mentions)
Lsm 510, by Leica (1 mentions)
Sp8 gated sted microscope, by Leica (1 mentions)
4 protocols using bx51 upright microscope
1
Quantifying Cardiac Fibrosis by Trichrome Staining
2
Microscopy Imaging Techniques
3
Automated Microscopy for Cerebellum
4
Fluorescence Imaging for Pyramidal Cell Density
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For fluorescence imaging, coronal slices (200 mm thick) were suspended in a solution of DAPI. Images were acquired on an Olympus BX51 upright microscope and a Leica SP8 gated STED microscope.
In order to estimate the density of expression, we carried out pyramidal cell count analysis in the cell-body layer near the injection site. First, we confirmed that nuclear DAPI staining corresponded to pyramidal neurons as detected in adjacent Nissl-stained brain sections. We estimated the number of cell bodies with clear presence of EYFP in a standardized cell-body zone of interest (400 mm long). Cell counts were performed three times, and an average for each panel was taken.
In order to estimate the density of expression, we carried out pyramidal cell count analysis in the cell-body layer near the injection site. First, we confirmed that nuclear DAPI staining corresponded to pyramidal neurons as detected in adjacent Nissl-stained brain sections. We estimated the number of cell bodies with clear presence of EYFP in a standardized cell-body zone of interest (400 mm long). Cell counts were performed three times, and an average for each panel was taken.
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