Each group of samples that were harvested at 2 and 12 weeks in vivo was fixed in 4% formaldehyde, dehydrated, and embedded in paraffin. Serial sections were obtained and subjected to H&E staining. Digital images with 20‐fold magnifications were captured using an image acquisition system (Olympus, Japan) and integrated into a panoramic view. Intact adipocytes that were distributed at the peripheral and central regions were counted (average from 10 high‐magnification fields per section), respectively.
Image acquisition system
Manufactured by Olympus
Sourced in Japan
The Image acquisition system is a versatile laboratory equipment designed to capture high-quality digital images. It enables the user to acquire, process, and analyze visual data from various samples or specimens.
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Lab products found in correlation
5 protocols using image acquisition system
1
In Vivo Adipocyte Morphology Assessment
2
Immunohistochemical Staining for CD31 Analysis
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For immunohistochemical (IHC) staining, the paraffin-embedded tissue samples were firstly sectioned to a thickness of 10 μm and mounted on glass slides. Then, these sections were deparaffinized in xylene and rehydrated through a graded ethanol series. After washing, the sections were blocked in 3% hydrogen peroxide and treated with sodium citrate buffer (10 mM sodium citrate, 0.05% tween 20, pH 6.0) for retrieval for 30 min at 95 °C. After blocking in 5% BSA for 1 h at room temperature, the sections were incubated with primary antibodies against CD31 (ab28364; Abcam) overnight at 4 °C. Next, the sections were incubated with HRP-conjugated secondary antibodies. Also, hematoxylin and eosin (H&E) stain was performed to observe neovascularization and tissue morphology. The digital images of these stained sections were taken using an image-acquisition system (Olympus) and were analyzed using Motic DSAssistant software (Motic VM 3.0).
3
Quantitative Histological Analysis of Metabolic Tissues
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Rats fed for 1, 2, 4, and 6 m were sacrificed, and liver, omentum fat and femurs were harvested and fixed with 4% paraformaldehyde (Sigma-Aldrich, St. Louis, MO, United States) followed by H&E staining. Images were captured using the image acquisition system (Olympus, Shinjuku, Japan). The percentage of fat vacuole area within hepatocytes in liver sections, diameters of adipocytes in omentum fat sections and numbers of osteoclasts in femur sections were measured separately by assessing 3 randomly selected high-power microscope fields in each slide (n = 5 per group).
4
CD34 Immunohistochemistry of Paraformaldehyde-Fixed Tissue
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On POD 7, samples from the CVZ which were fixed in paraformaldehyde were sectioned into 5 μm slices. Sections were rehydrated in a graded series of ethanol after they were deparaffinized through xylene. Then, sections were immersed in 3% H2O2 and incubated to saturate nonspecific sites. Last sections were incubated with CD34 (1:50; Abcam, Cambridge, UK) at 4 °C overnight. Sections were imaged at × 100 magnification on an image acquisition system (Olympus, Tokyo, Japan). The number of CD34-positive microvessels was calculated in five dense fields.
5
Immunohistochemical Analysis of METTL3
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IHC staining was performed as described previously (46 (link)). Briefly, TMAs were treated with xylene and 100% ethanol, followed by decreasing concentrations of ethanol. After antigen retrieval, TMAs were blocked and stained with anti-METTL3 antibody (1:500, Abcam, USA), followed by incubation with secondary antibody and standard avidin biotinylated peroxidase complex. Hematoxylin was used for counterstaining, and images were obtained with an Image acquisition system (Olympus, Japan). The total METTL3 immunostaining score was calculated as the sum of the score for the proportion of positively stained tumor cells (PP) and the score for staining intensity (SI). PP was scored with a four-point scale: 0 (< 5%), 1 (5–25%), 2 (25–50%), 3 (50%-75%), and 4 (>75%), and SI was scored on a scale of 0 to 3 (0, negative staining; 1, weak staining; 2, moderate staining; and 3, strong staining). The final staining score was calculated by multiplying the SI and PP scores, resulting in a score value ranging from 0 to 12. The median value of the total staining score was 4; thus, a score of 0-4 was defined as low expression, and a score of 4-12 was defined as high expression.
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