Neutral lipid accumulation was visualized using fluorescent dye staining, Nile red, in live fish as previously described (25 (link), 38 (link)). Nile red (N3013; Sigma) was dissolved to a concentration of 0.1 g/mL. The fish were immersed in Nile red at 28.5°C overnight in the dark. Images were taken using an Olympus SZX16 FL stereomicroscope (Olympus, Tokyo, Japan) at an excitation wavelength of 488 nm. Liver triglycerides were measured using commercially available kits (A110-1, Nanjing Jiancheng Bioengineering Institute, China). Fatty droplet accumulation in the liver was visualized using Oil Red O staining as previously described (25 (link)).
Szx16 fl stereomicroscope
Manufactured by Olympus
Sourced in Japan
The Olympus SZX16 FL stereomicroscope is a high-performance optical instrument designed for a variety of laboratory applications. It features a zoom ratio of 16:1, providing a wide range of magnification capabilities. The microscope is equipped with an LED illumination system and is capable of fluorescence imaging.
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Lab products found in correlation
3 protocols using szx16 fl stereomicroscope
1
Visualization and Quantification of Lipid Accumulation
2
Quantifying Zebrafish Lipid Metabolism
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Visceral mass and muscle TG were measured using a Triglyceride Assay Kit (A110-1-1, Nanjing Jiancheng Bioengineering Institute Co., Ltd., China) following the manufacturer’s instructions. Neutral lipid staining was performed using Nile Red dye (N3013, Sigma, USA) at a working concentration of 0.1 μg/mL for 12 h in the dark, followed by acquisition of fluorescent images using an Olympus SZX16FL stereomicroscope (Olympus, Japan) at an excitation wavelength of 488 nm (Xi et al., 2023 (link); Yang et al., 2018 (link)). Total lipid content (percent dry weight) was measured using the Folch method (Folch et al., 1957 (link)). Briefly, individual vacuum freeze-dried zebrafish were cut up in 5 mL of chloroform (Sinopharm Chemical Reagent Co., Ltd., China):methanol (Sinopharm Chemical Reagent Co., Ltd., China) (2:1, vol/vol), extracted for 2 h, and centrifuged at 750 ×g for 5 min at room temperature. The upper phase was then vortexed with 0.4% calcium chloride (Sinopharm Chemical Reagent Co., Ltd., China), with the upper water phase discarded and the lower organic phase dried and weighed to measure total lipid mass per fish.
3
Nile Red Staining of Zebrafish
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Nile Red (N3013; Sigma Aldrich, St. Louis, MO, USA) was dissolved in acetone at 1 mg/mL as the stock solution. Mutants and size-matched control zebrafish were immersed in system water containing 0.1 μg/mL Nile Red for 1 hr for larval fish. Images were obtained using an Olympus SZX16 FL Stereo Microscope (Olympus, Tokyo, Japan) at an excitation wavelength of 488 nm.
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