Ten μm cryosections of the aortic root, serially collected starting from the tricuspid valves, were stained with Oil red O to identify atherosclerotic lesions. Mean lesion area from 5 sections per mouse was quantified by using a Leica image analysis system, consisting of a Leica DMRE microscope coupled to a camera and Leica QWin Imaging software (Leica Ltd., Cambridge, UK). Lesional collagen content was determined from Masson’s Trichrome Accustain (Sigma-Aldrich)-stained sections. Lesion analyses were performed in a blinded manner.
Qwin imaging software
Manufactured by Leica
Sourced in United Kingdom
Leica Qwin is an imaging software that provides digital image analysis and processing capabilities. It is designed to be used with Leica microscopes and digital cameras to capture, analyze, and manage microscopic images.
Automatically generated - may contain errors
Lab products found in correlation
Dmre microscope, by Leica (5 mentions)
Cm3050s cryostat, by Leica (2 mentions)
Tissue tek oct compound, by Sakura Finetek (2 mentions)
Masson s trichrome accustain, by Merck Group (1 mentions)
Masson s trichrome, by Merck Group (1 mentions)
Oil red o staining, by Merck Group (1 mentions)
α smooth muscle actin, by Merck Group (1 mentions)
Tunel kit, by Roche (1 mentions)
5 protocols using qwin imaging software
1
Quantification of Aortic Atherosclerosis
2
Aortic Root Histomorphometric Analysis
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Aortic roots were sectioned transversely at 10 µm intervals using a CM3050 S cryostat (Leica Ltd., Rotterdam, The Netherlands) by embedding the hearts in Tissue-Tek O.C.T. Compound (Sakura Finetek, Alphen aan den Rijn, NL). A Leica DMRE microscope and Leica Qwin Imaging software (Leica Ltd., Cambridge, UK) were used to obtain images. Atherosclerotic lesion areas, total vessel areas, and lesional lipid, macrophage, and collagen contents were quantified in a blinded manner. Neutral lipids were identified using a standard Oil red O staining (Sigma, Steinheim, Germany). Masson’s Trichrome (Sigma, Steinheim, Germany) was used to stain collagen fibers with hematoxylin as the counterstain. Macrophages were stained immunohistochemically using an anti-CD68 antibody. Cholesterol crystals were visualized under polarized light.
3
Quantifying Aortic Lesions in Mcl-1 Mice
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Two hours before sacrifice, Mcl-1−/− or WT mice (n = 5) received intraperitoneal injections of CXCL1 (200 ng/ml in 1 ml PBS) or PBS control. The mice were anesthetized and perfused with PBS. Cryosections of the aortic root tissue were stained with hematoxylin and eosin (HE) or Oil Red O. Lesion size was quantified using a Leica DMRE microscope with camera and Leica Qwin Imaging software (Leica Ltd). MGCs were defined as macrophages with two or more round nuclei on the HE slides and quantified by an animal pathologist. Immunohistochemical stainings were performed for macrophage (MOMA-2, Sigma) and vSMC (α-smooth muscle actin, Sigma) content. Apoptotic cell content was quantified using terminal deoxytransferase dUTP nick-end labeling (TUNEL) kit (Roche Diagnostics).
4
Quantification of Atherosclerotic Lesions
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The mean atherosclerotic lesion area of each mouse was quantified taking the guidelines for experimental studies described in the AHA statement 22 (link) into account. Ten Oil red O-stained cryosections (10 µm), starting at the appearance of the tricuspid valves up to 300 µm of the ascending aorta, were analyzed using a Leica image analysis system, consisting of a Leica DMRE microscope coupled to a video camera and Leica Qwin Imaging software (Leica Ltd, Cambridge, United Kingdom).
5
Quantitative Liver Lipid Analysis
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Formalin‐fixed liver specimens were embedded in Tissue‐Tek® O.C.T. Compound (Sakura Finetek Europe, Alphen an den Rijn, NL), and cryosections (8 μm) were prepared on a Leica CM3050‐S cryostat (Leica Ltd., Cambridge, UK). Liver cryosections were stained for neutral lipids with Oil red O. Images were obtained with a Leica image analysis system, which consisted of a Leica DMRE microscope coupled to a video camera and Leica Qwin Imaging software (Leica Ltd., Cambridge, UK). For each liver, three images were obtained from three different liver sections per mouse. Images were blinded, and each image was assigned a score between 0 and 3 by an unbiased observer for the amount of red staining in the liver. Representative photomicrographs were taken for each group using the same system with 40× magnification.
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