Images of the wounds were captured with a 10-megapixel digital camera (Sony Corporation, Tokyo, Japan) at 7, 14 and 21 days, and an image analyzer (Image-Pro Plus 6.0; Media Cybernetics, Inc., Rockville, MD, USA) was used to obtain wound-size measurements. At the end of 7, 14 and 21 days, wound healing was quantified by calculating the remaining wound area for each group. The percentage of wound closure was calculated as follows: wound closure (%)=(area of original wound-area of actual wound)/area of original wound ×100. To assess the wound histologically, mice at day 14 were sacrificed by cervical dislocation for histological assessment. The harvested wound areas, including a border of normal tissue, were immediately fixed in 10% neutral-buffered formalin. The specimens were embedded in paraffin, sectioned into 5 µm slices, and stained with hematoxylin and eosin to determine the quality of wound healing using a Nikon E200® photomicroscope equipped with the Moticam 2300® image capture system (10 (link)). The scalded tissue samples at 7 days were collected and maintained at −80°C for further evaluation.
Moticam 2300
Manufactured by Nikon
Sourced in United States
The Moticam 2300 is a digital microscope camera designed for laboratory use. It captures high-quality images and video with a resolution of up to 3.0 megapixels. The camera is compatible with a variety of microscopes and can be connected to a computer or display device for image viewing and analysis.
Automatically generated - may contain errors
3 protocols using moticam 2300
1
Wound Healing Quantification and Histological Assessment
2
Iron Detection in Labeled Cells
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Prussian blue staining was performed to test the presence of iron in the labeled cells using an Iron Stain Kit (Sigma-Aldrich), according to the supplier’s protocol. Staining results were examined by light microscopy and photomicrographs were taken using a Moticam 2300 camera mounted on a Nikon Diaphot microscope with Mtic Images Plus 2.0 software.
3
Quantifying Immunostained Cell Density
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For sections subjected to immunohistochemistry, images were acquired using a Moticam 2300 digital camera coupled to a Nikon Eclipse E200 microscope (Nikon, NY, USA) and MOTIC IMAGES PLUS 2.0 software. Images (1024 9 768 pixels) were acquired from 10 randomly selected fields of view for each slide, at 400 9 magnification without any knowledge of the grade of injury. The sizes of features in the images were determined with IMA-GEJ software 1.47 (National Institutes of Health, USA) using the freehand line tool. Images that were 1024 9 768 pixels corresponded to 170Á629 127Á97 lm. Using the colour deconvolution plug in of IMAGEJ, the separation of stains was achieved as described by Ruifrok and Johnston. 23 Using the cell counter plugin of IMAGEJ software, the number of immunostained cells was determined for each image, providing the average number of immunostained cells per mm 2 .
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