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PC6S is a small compound that emits strong fluorescence in lipid droplets. PC6S can dye clearly not only cellular lipid droplets but also tissue lipid droplets. Because PC6S has excellent selectivity, it can be used in combination with multiple fluorescent dyes. PC6S emits green fluorescence (λex, max = 468 nm, λem, max = 535 nm, in dibutyl ether).
Application: multiple staining of cells
Figure. A : PC6S (Lipid droplet), B : Hoechst® 33342 (Nucleus), C : MitoTracker® Red (Mitochondria), D : Merged image, Each scale bar is 20 μm.
‡These figures are provided by Dr. Toshitada Yoshihara.
Procedure
- Add oleic acid to medium of HeLa cells and incubate for 24 hours.
- Replace with medium containing PC6S, Hoechst® 33342, MitoTracker® Red and incubate at 37 °C for 30 minutes.
- Remove the medium and wash twice with PBS(-).
- Observe through fluorescence microscope.
Hoechst® is a registered trademark of Hoechst GmbH. MitoTracker® is a registered trademark of Molecular Probes, Inc.
Reference
- Visualization of Lipid Droplets in Living Cells and Fatty Livers of Mice Based on the Fluorescence of π-Extended Coumarin Using Fluorescence Lifetime Imaging Microscopy
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Example of staining with Oil Red O
Oil red O is a diazo dye which has been widely used for staining fat cell and lipids since long ago. This lysochrome (fat-soluble dye) red dye can be used for staining of neutral triglycerides and low polar lipids. Oil Red O staining is done on fresh or frozen samples, since alcohol fixation removes lipid. Its usage is simple, and requires only washing after adding the staining solution, which makes it easy to identify lipids visually.
Furthermore, accumulation of lipids can be quan-tified by eluting the dye using isopropanol after staining and measuring the absorbance.
Figure. 10 days later of addition of media for differentiation to 3T3-L1 cell and then stained by 1mg/mL of O0483.