Morphometric analyses were based on techniques described elsewhere55 . Briefly, hearts and kidneys were dehydrated, cleared, and embedded in paraffin. Paraffin blocks were cut into 4–5-μm thick sections, and adjacent sections were stained with haematoxylin/eosin for evaluation of general renal and myocardial morphology and identification of possible tissue damage. All morphometric measurements were made in tissue sections under a light microscope (Leica DM5000) and analysed using the Leica Qwin Image Processing and Analysis Software (Germany). To assess high-sodium diet effect on heart, the left ventricle Wt and left ventricle lumen diameter (LVL) were determined on sections at 1x magnification56 (link). Cuts (3 to 5 sections) were performed at the same level of the hearts, transversally and perpendicularly with the longitudinal axis, at the middle level of the ventricles where the ventricles had bigger diameter. The degree of cardiac hypertrophy was calculated as the ratio between left ventricle Wt and LVL (see supplementary data). In order to evaluate the extent of renal damage imposed by the high-sodium diet, glomerulosclerosis was assessed by counting the number of sclerotic glomeruli and express it as the percent of total glomeruli counted in a sequence of 25 field and by the ratio between glomerular tuft area (GTA) and Bowman’s capsule area (BCA)56 (link).
Qwin image processing and analysis software
Manufactured by Leica
Sourced in Germany
Qwin Image Processing and Analysis Software is a digital imaging application designed for the analysis and processing of microscope images. It provides a set of tools for image acquisition, enhancement, measurement, and quantification. The software is compatible with a wide range of microscope systems and supports various image file formats.
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Lab products found in correlation
5 protocols using qwin image processing and analysis software
1
Morphometric Analysis of Cardiac and Renal Tissue
2
Quantifying Lung Morphometry via Image Analysis
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For mean linear intercept (MLI) calculations, hematoxylin-eosin-stained paraffin sections were imaged with a bright field Axioimager microscope (Zeiss, Oberkochen, Germany) at 40X magnification. Measurements of average wall thickness, airspace percentage, and mean linear intercept were made with Leica Qwin Image Processing and Analysis Software.
3
Lung Histology and Morphometry
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Lungs were immersed in 10% phosphate buffered formalin for 24 h and kept in 70% ethanol until embedding in paraffin. Tissues were sliced (5-μm sections) and stained with hematoxylin/eosin for light microscopy examination. Morphometrical analysis was performed using a Leica DM 5000B digital camera and Leica Q Win Image Processing and Analysis Software. For each different staining, the area of positivity was measured in mm2 for 3 bronchioles per slide.
4
Histopathological Analysis of Cardiac and Renal Tissues
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For the histopathological analysis, hearts and kidneys were collected and fixed in
10% neutral-buffered formalin solution. After 72 h of fixation, the hearts and
kidneys were dehydrated, cleared, and embedded in paraffin. The paraffin block was
cut into 4- to 5-μm-thick sections, and adjacent sections were stained with either
hematoxylin/eosin for evaluation of general myocardial and renal damage or Masson's
trichrome for quantification of collagen-tissue deposition. Morphometric evaluations
were made in tissue sections under an optical microscope (DM5000; Leica, Germany) and
analyzed with QWin Image Processing and Analysis Software (Leica) in 20 optical
microscope images at 40× magnification for each animal. In the hearts, the
cardiomyocyte diameter was measured by a previously described method (2 (link)) in 20 optical microscope images at 40×
magnification. The left ventricle wall thickness (Wt) and ventricle lumen (L) were
measured on sections at 5× magnification, and the degree of cardiac hypertrophy was
calculated as the Wt/L ratio. Higher Wt/L ratios indicated concentric hypertrophy,
and lower Wt/L ratios indicated eccentric hypertrophy. Because the SHAM SED rats did
not show changes in these ratios, the Wt/L ratio of these animals was used as a
control. The cardiac and renal inflammatory process and tissue collagen deposition
were also quantified as previously described (2 (link)).
10% neutral-buffered formalin solution. After 72 h of fixation, the hearts and
kidneys were dehydrated, cleared, and embedded in paraffin. The paraffin block was
cut into 4- to 5-μm-thick sections, and adjacent sections were stained with either
hematoxylin/eosin for evaluation of general myocardial and renal damage or Masson's
trichrome for quantification of collagen-tissue deposition. Morphometric evaluations
were made in tissue sections under an optical microscope (DM5000; Leica, Germany) and
analyzed with QWin Image Processing and Analysis Software (Leica) in 20 optical
microscope images at 40× magnification for each animal. In the hearts, the
cardiomyocyte diameter was measured by a previously described method (2 (link)) in 20 optical microscope images at 40×
magnification. The left ventricle wall thickness (Wt) and ventricle lumen (L) were
measured on sections at 5× magnification, and the degree of cardiac hypertrophy was
calculated as the Wt/L ratio. Higher Wt/L ratios indicated concentric hypertrophy,
and lower Wt/L ratios indicated eccentric hypertrophy. Because the SHAM SED rats did
not show changes in these ratios, the Wt/L ratio of these animals was used as a
control. The cardiac and renal inflammatory process and tissue collagen deposition
were also quantified as previously described (2 (link)).
5
Atherosclerosis Pathological Evaluation
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At the end of 12 weeks, arteries were collected from the rats to perform HE staining and observe pathological changes under a light microscope and electron microscope. A Zeiss-Axioskop 20 microscope was used to observe the histological changes in HE-stained sections, and an Axiocan HRc camera was employed to obtain micrographs. Leica Qwin Image Processing and Analysis Software was used to analyze AS lesions and the cross-sectional area of the artery lumen and to calculate the relative area of atherosclerosis lesions (the ratio of the lesion area versus the lumen cross-sectional area), which is represented as a percentage (%). Blood lipids, liver lipids, blood rheology, inflammatory factors, and the gene expression and protein expression of eNOS, iNOS, IKKβ, p-IKKβ, and NF-κBp65 in artery tissue were examined.
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