Protocol detail

Lipid Droplet Quantification in Cultured Cells

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Cultured cells were fixed with 4% paraformaldehyde (PFA) and 2% sucrose and then stained with Nile red (Sigma-Aldrich, St. Louis, MO) and High-Content Screening (HCS) Cell Mask Blue (Invitrogen, Waltham, MA) according to the manufacturer’s protocols. Images were acquired using the Opera high-content screening system (20 x confocal lens) and lipid droplets intensity per cell was determined using the Columbus Image Analysis System (Perkin Elmer, Waltham, MA) (Liu et al., 2020 (link)).

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Ge M., Molina J., Kim J.J., Mallela S.K., Ahmad A., Varona Santos J., Al-Ali H., Mitrofanova A., Sharma K., Fontanesi F., Merscher S, & Fornoni A. (2023). Empagliflozin reduces podocyte lipotoxicity in experimental Alport syndrome. eLife, 12, e83353.

Publication 2023

Nile red Columbus Image Analysis System

Cell Cultured cells Lens Lipid droplets Paraformaldehyde Sucrose

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Lipid droplets intensity per cell

control variables

Cells were fixed with 4% paraformaldehyde (PFA) and 2% sucrose
Cells were stained with Nile red and High-Content Screening (HCS) Cell Mask Blue according to the manufacturer's protocols

controls

No positive or negative controls were explicitly mentioned in the provided information.

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