Epididymal and inguinal subcutaneous WAT was fixed in 10% formalin neutral buffer (FUJIFILM Wako Pure Chemical Corporation, Tokyo, Japan) for 24 h at 4 °C and embedded in paraffin following the standard protocol. Sections (5 μm) were stained with hematoxylin and eosin for 10 and 5 min at room temperature, respectively, and at least 750 adipocytes in eWAT from each mouse were visualized using a BZ-X810 microscope (Keyence, Osaka, Japan) and enumerated using a hybrid cell count software (BZ-X800 Analyzer, ver. 1.1.2.4, Keyence).
Formalin neutral buffer
Manufactured by Fujifilm
Sourced in Japan
Formalin neutral buffer is a laboratory reagent used to prepare and preserve tissue samples. It maintains a neutral pH to ensure proper fixation and stabilization of biological specimens for downstream analysis.
Automatically generated - may contain errors
Lab products found in correlation
Bz x810 microscope, by Keyence (1 mentions)
Bz x800 analyzer, by Keyence (1 mentions)
Eosin solution, by Fujifilm (1 mentions)
Hematoxylin solution, by Fujifilm (1 mentions)
Liquid nitrogen cooled isopentane, by Fujifilm (1 mentions)
Bz x analyzer software, by Keyence (1 mentions)
Cryostat, by Leica Microsystems (1 mentions)
Rnalater solution, by Qiagen (1 mentions)
Pentobarbital sodium, by Sumitomo Pharma (1 mentions)
3 protocols using formalin neutral buffer
1
Adipocyte quantification in white adipose tissue
2
Histological Evaluation of Muscle Fibers
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The gastrocnemius muscles were isolated for hematoxylin and eosin (H&E) staining. The muscles were frozen in a liquid nitrogen-cooled isopentane (Wako Chemicals). Subsequently, ten or sixteen-micrometer thick cross sections were cut from the middle portion of the muscle using a cryostat (Leica Microsystems). Sections were fixed in 10% formalin neutral buffer (Wako Chemicals) and then stained with hematoxylin solution (Wako Chemicals) followed by the eosin solution (Wako Chemicals). Cross-sectional area (CSA) of muscle fibers was determined using BZ-X analyzer software (Keyence). >500 fibers per mouse were analyzed from three areas in each muscle.
3
Femur, Muscle Tissue Harvesting Protocol
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Rats were euthanized by an injection of sodium pentobarbital (150 mg/kg body weight) (Dainippon Sumitomo Pharma Co. Ltd., Osaka, Japan), and the right femur, left tibialis anterior muscle, and soleus muscle were harvested for evaluations after 2 or 6 weeks of treatment.
The right femur was dipped in 10% formalin neutral buffer (Wako Pure Chemical Industries, Osaka, Japan) for BMD measurement. The left tibialis anterior muscle was immediately frozen in liquid nitrogen and stored at -80°C for histological analyses. The soleus muscle was stored in RNAlater solution (Qiagen, Hilden, Germany) at -80°C for gene expression analysis.
The right femur was dipped in 10% formalin neutral buffer (Wako Pure Chemical Industries, Osaka, Japan) for BMD measurement. The left tibialis anterior muscle was immediately frozen in liquid nitrogen and stored at -80°C for histological analyses. The soleus muscle was stored in RNAlater solution (Qiagen, Hilden, Germany) at -80°C for gene expression analysis.
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