Ventricle samples were fixed in 4% paraformaldehyde and paraffin embedded for histological analysis at 6 µm cross-sections. Cardiac collagen deposition/interstitial fibrosis was assessed by Masson’s trichrome stain (Alfred Pathology, Melbourne, Australia). Images of the LV were obtained using an Olympus light microscope at 40x magnification. Collagen stained blue, which was measured and analyzed using Olympus Image-Pro Plus version 6.0. The percentage fibrosis was calculated by dividing the total area of collagen by the total area of the LV and multiplying by 100%. Data were normalized to a control value of 1 and presented as a fold change.
Image pro plus version 6
Manufactured by Olympus
Sourced in Japan
Image-Pro Plus Version 6.0 is an image analysis software developed by Olympus. It provides tools for capturing, processing, measuring, and analyzing digital images. The software supports a wide range of image file formats and offers features for image enhancement, segmentation, and object recognition.
Automatically generated - may contain errors
4 protocols using image pro plus version 6
1
Quantifying Cardiac Fibrosis via Histology
2
Quantifying Cardiac Fibrosis via Histology
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Ventricle samples were fixed in 4% paraformaldehyde and paraffin embedded for histological analysis and cut at 6 μm cross-sections. Masson’s trichrome stain was performed to assess cardiac collagen deposition/interstitial fibrosis (Alfred Pathology, Melbourne, Australia). An Olympus light microscope at 40x magnification was employed to capture images of the LV and the Olympus Image-Pro Plus Version 6.0 was used to enumerate collagen stained blue. To calculate the proportion of fibrosis, the total area of collagen was divided by the total area of the LV and then multiplied by 100%. Data were normalized to a control value of 1 and presented as a fold change.
3
Histological Analysis of Skin Samples
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Skin samples were fixed in 10% formalin, dehydrated, embedded in paraffin, and cut into sections 5 µm thick. After deparaffinizing in xylene and rehydrating using an alcohol series, the sections were stained with hematoxylin and eosin (H&E), Masson’s trichrome, and Picrosirius red (Solarbio, Beijing, China). Images were randomly photographed with an upright transmission fluorescence microscope (Olympus, Tokyo, Japan) and analyzed by Image-Pro Plus Version 6.0.
4
Neurological Scoring and Infarct Volume Analysis
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Twenty‐four hours after I/R a 4‐tiered neurological scoring system and infarct volume were used to determine the outcome by a blinded observer as described previously.16 Postural reflex was scored on a four‐point grade scale: 0, normal function; 1, flexion of the torso and contralateral forelimb on lifting the animal by the tail; 2, circling to the contralateral side but normal posture at rest; 3, reclination to the contralateral side at rest; and 4, absence of spontaneous motor activity. TTC infarct measurement techniques were performed to measure infarct size. The brains were sliced into 2 mm thick coronal sections for staining with 2% TTC in phosphate buffer saline at 37°C for 20 minutes. Infarction volume was measured by digital imaging (Digital Camera, Olympus MDF‐382E) and image analysis software (Image‐Pro Plus Version 6.0). The infarct area was calculated across each section and was presented as a percentage relative to the area of the contralateral hemisphere.
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