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Deformation of lipid droplets in fixed samples
Satoshi Fukumoto1, Toyoshi Fujimoto
1Department of Anatomy and Molecular Cell Biology, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan.
Histochemistry and Cell Biology
|November 15, 2002
Summary
Nile red is a superior stain for observing lipid droplet structure in fluorescence microscopy. Unlike Sudan III and oil red O, it avoids artifactual fusion caused by ethanol, isopropanol, and glycerol.
Area of Science:
- Cell Biology
- Microscopy Techniques
Background:
- Lipid droplets (LDs) are crucial cellular components.
- Accurate visualization of LDs is essential for understanding their function.
Purpose of the Study:
- To investigate the differing appearances of LDs stained with Nile red versus Sudan III and oil red O.
- To identify factors causing artifactual changes in LD morphology during staining.
Main Methods:
- Sequential staining and photography of LDs using Nile red, Sudan III, and oil red O.
- Evaluation of staining artifacts induced by ethanol, isopropanol, and glycerol.
- Comparative immunofluorescence labeling for adipose differentiation-related protein (ADRP).
Main Results:
- Ethanol, isopropanol, and glycerol caused fusion of adjacent LDs, even in fixed samples.
- This fusion artifact resulted in ADRP dislocation to the periphery of enlarged LDs.
- Nile red staining revealed LD structure without inducing fusion artifacts.
Conclusions:
- Nile red provides a more accurate representation of native lipid droplet structure compared to Sudan III and oil red O.
- Solvents and mounting media commonly used in lipid staining can induce significant morphological artifacts.