Frozen liver sections were stained with Oil Red solution (Servicebio, Wuhan, China) for 8–10 min in the dark. The sections were immersed in 60% isopropanol for differentiation, followed by incubation with hematoxylin and bluing in tap water. The red-stained lipid droplets were observed via light microscopy. Quantification of the Oil Red O intensity (Oil Red O staining area per cell) was performed using ImagePro Plus software (Media Cybernetics, Georgia, USA).
Oil red solution
Manufactured by Wuhan Servicebio Technology
Sourced in China
Oil Red solution is a lipid-soluble dye used for the histological staining of neutral lipids and lipoproteins. It is commonly utilized in various laboratory applications, such as the identification and visualization of fat cells and lipid droplets in tissue sections.
Automatically generated - may contain errors
Lab products found in correlation
Hematoxylin, by Wuhan Servicebio Technology (2 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
Dm4000 b led, by Leica (1 mentions)
Eclipse e100, by Nikon (1 mentions)
Eclipse e100 microscope, by Nikon (1 mentions)
Sirius red solution, by Wuhan Servicebio Technology (1 mentions)
Vs200, by Olympus (1 mentions)
Safranino fast green staining solution, by Wuhan Servicebio Technology (1 mentions)
G1015, by Wuhan Servicebio Technology (1 mentions)
5 protocols using oil red solution
1
Quantification of Hepatic Lipid Droplets
2
Histopathological Analysis of Liver
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Liver samples were secured in paraformaldehyde (4%) and addressed for dehydration, paraffin embedding, and were sectioned (4 μm). The sections were then deparaffinized and stained with hematoxylin and eosin. An upright optical microscope (Nikon, NIKON ECLIPSE E100) was employed to observe the results.
Frozen liver sections, which were stained with Oil Red solution (Servicebio G1016) for 10 min and immersed in hematoxylin (Servicebio G1004) for counterstaining for 5 min, were observed under a microscope (LEICA DM 4000 B LED). The lipid accumulation area was analyzed by Image-Pro Plus 6.0 by evaluating integrated optical density (IOD).
The above staining procedure referred to the reported method with some modification [23 (link)]. The histological evaluation of liver slices was random.
Frozen liver sections, which were stained with Oil Red solution (Servicebio G1016) for 10 min and immersed in hematoxylin (Servicebio G1004) for counterstaining for 5 min, were observed under a microscope (LEICA DM 4000 B LED). The lipid accumulation area was analyzed by Image-Pro Plus 6.0 by evaluating integrated optical density (IOD).
The above staining procedure referred to the reported method with some modification [23 (link)]. The histological evaluation of liver slices was random.
3
Lipid Quantification in Tissue Sections
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The slices were reheated and fixed in the fixative solution for 15 min. Next, the slices were stained with Oil Red solution (Servicebio, China) for 10 min in the dark. Then, the slices were immersed in two cup of 60% isopropanol for differentiation in turn, 3 s and 5 s respectively. Subsequently, the slices were re-stained with hematoxylin (Servicebio, China) for 1 min and rinsed with running water. Finally, the slices were sealed with glycerin gelatin and observed under a Nikon ECLIPSE E100 microscope. The individuals who quantified the microscopy were blinded.
4
Histological Assessment of Hepatic Steatosis and Fibrosis
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At the end of treatment, histological analysis was performed to evaluate hepatocellular steatosis and fibrosis. For hematoxylin-eosin (H&E) staining, paraffin-embedded liver sections were incubated with H&E solutions (Servicebio, Wuhan, China). Paraffin-embedded sections were also stained with Sirius Red solution (Servicebio, Wuhan, China). Subsequently, sections were dehydrated with anhydrous ethanol and xylene and sealed with neutral gum. Frozen liver tissue slices were incubated with Oil Red solution (Servicebio, Wuhan, China) in the dark and immersed in hematoxylin for 3–5 min. The slices were then sealed with glycerin-gelatin. Histological assessment and scoring were performed by a pathologist blinded to the study. The NAFLD Activity Scoring (NAS) (steatosis/inflammation/ballooning) were performed using the clinical criteria outlined by Kleiner et al.28 (link) Using Image J software to quantify the percentage of lipid droplet area and fibrosis positive area of the entire image area.
5
Paraffin Embedding and Staining of Plant Cuticle
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The fresh fruits were trimmed from the target tissue with a scalpel and put into fixative (G1101, Wuhan Servicebio Technology Co., Ltd, China) for more than 24 h. After dehydration with gradient alcohol, the tissue is embedded in paraffin wax at 65 °C. The slice transverse sections of 4 μm thickness were cut with the paraffin slicer and stained with Safranin O‐Fast Green staining solution (G1031, Servicebio). Oil‐Red solution (G1015, Servicebio) stained slices were chosen for cuticle observation. Multiplex slice images were acquired through a whole slide imaging system (Olympus VS200) at 20× objective. Cytological assessment, including cuticle thickness, cell size and cell number, was performed using ImageJ software.
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