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Lipid droplet quantification using the Columbus™ software. (a) Human podocytes nucleus stained with DAPI (Blue), lipid droplets with BODIPY 493/503 (green) and cytosol with CellMask Deep Red (Red); (b) identification of cell nuclei; (c) identification of the cell cytoplasm; (d) exclusion of the cells at the image border. Cells marked with green were selected for further analysis, and cells marked with red were removed from the analysis; (e) spot detection. Detected spots are marked with white. (f) LDL treatment increases LDs in cultured podocytes. Human podocytes were incubated without (control) or with 50 μg/ml acetylated low-density lipoprotein (LDL) for 24 h at 37 °C. Cells were fixed and stained as described in protocol. LDs were quantified using Columbus software
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Lipid droplet quantification using the Columbus™ software. (a) Human podocytes nucleus stained with DAPI (Blue), lipid droplets with BODIPY 493/503 (green) and cytosol with CellMask Deep Red (Red); (b) identification of cell nuclei; (c) identification of the cell cytoplasm; (d) exclusion of the cells at the image border. Cells marked with green were selected for further analysis, and cells marked with red were removed from the analysis; (e) spot detection. Detected spots are marked with white. (f) LDL treatment increases LDs in cultured podocytes. Human podocytes were incubated without (control) or with 50 μg/ml acetylated low-density lipoprotein (LDL) for 24 h at 37 °C. Cells were fixed and stained as described in protocol. LDs were quantified using Columbus software
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Lipid droplet quantification using the Columbus™ software. (a) Human podocytes nucleus stained with DAPI (Blue), lipid droplets with BODIPY 493/503 (green) and cytosol with CellMask Deep Red (Red); (b) identification of cell nuclei; (c) identification of the cell cytoplasm; (d) exclusion of the cells at the image border. Cells marked with green were selected for further analysis, and cells marked with red were removed from the analysis; (e) spot detection. Detected spots are marked with white. (f) LDL treatment increases LDs in cultured podocytes. Human podocytes were incubated without (control) or with 50 μg/ml acetylated low-density lipoprotein (LDL) for 24 h at 37 °C. Cells were fixed and stained as described in protocol. LDs were quantified using Columbus software
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Lipid droplet quantification using the Columbus™ software. (a) Human podocytes nucleus stained with DAPI (Blue), lipid droplets with BODIPY 493/503 (green) and cytosol with CellMask Deep Red (Red); (b) identification of cell nuclei; (c) identification of the cell cytoplasm; (d) exclusion of the cells at the image border. Cells marked with green were selected for further analysis, and cells marked with red were removed from the analysis; (e) spot detection. Detected spots are marked with white. (f) LDL treatment increases LDs in cultured podocytes. Human podocytes were incubated without (control) or with 50 μg/ml acetylated low-density lipoprotein (LDL) for 24 h at 37 °C. Cells were fixed and stained as described in protocol. LDs were quantified using Columbus software
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Lipid droplet quantification using the Columbus™ software. (a) Human podocytes nucleus stained with DAPI (Blue), lipid droplets with BODIPY 493/503 (green) and cytosol with CellMask Deep Red (Red); (b) identification of cell nuclei; (c) identification of the cell cytoplasm; (d) exclusion of the cells at the image border. Cells marked with green were selected for further analysis, and cells marked with red were removed from the analysis; (e) spot detection. Detected spots are marked with white. (f) LDL treatment increases LDs in cultured podocytes. Human podocytes were incubated without (control) or with 50 μg/ml acetylated low-density lipoprotein (LDL) for 24 h at 37 °C. Cells were fixed and stained as described in protocol. LDs were quantified using Columbus software

Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Detection and Quantification of Lipid Droplets in Differentiated Human Podocytes

doi: 10.1007/978-1-4939-9488-5_17

Figure Lengend Snippet: Lipid droplet quantification using the Columbus™ software. (a) Human podocytes nucleus stained with DAPI (Blue), lipid droplets with BODIPY 493/503 (green) and cytosol with CellMask Deep Red (Red); (b) identification of cell nuclei; (c) identification of the cell cytoplasm; (d) exclusion of the cells at the image border. Cells marked with green were selected for further analysis, and cells marked with red were removed from the analysis; (e) spot detection. Detected spots are marked with white. (f) LDL treatment increases LDs in cultured podocytes. Human podocytes were incubated without (control) or with 50 μg/ml acetylated low-density lipoprotein (LDL) for 24 h at 37 °C. Cells were fixed and stained as described in protocol. LDs were quantified using Columbus software

Article Snippet: BODIPY 493/503 stock solution (Invitrogen, 1 mg/ml in 100% ethanol).

Techniques: Software, Staining, Cell Culture, Incubation