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A pitfall in using BODIPY dyes to label lipid droplets for fluorescence microscopy
Yuki Ohsaki1, Yuki Shinohara, Michitaka Suzuki
1Department of Anatomy and Molecular Cell Biology, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa, Nagoya, 466-8550, Japan.
Histochemistry and Cell Biology
|March 2, 2010
Summary
BODIPY 493/503 dye, used for lipid droplet (LD) imaging, can unexpectedly emit red fluorescence. This protocol prevents misinterpretation during cell biology studies of LDs.
Area of Science:
- Cell Biology
- Biochemistry
- Microscopy
Background:
- Lipid droplets (LDs) are crucial cellular components increasingly studied using fluorescence microscopy.
- BODIPY 493/503 is a widely used lipophilic dye for LD labeling, known for its green fluorescence.
- The dye's utility in double labeling experiments is recognized, but potential spectral shifts are a concern.
Purpose of the Study:
- To address the potential for BODIPY 493/503 to emit red fluorescence under specific conditions.
- To prevent erroneous interpretations in cell biological analyses of lipid droplets.
- To propose a reliable protocol for accurate LD fluorescence labeling.
Main Methods:
- Investigated the spectral properties of BODIPY 493/503 under various experimental conditions.
- Developed and validated a protocol to mitigate unexpected red fluorescence emission.
- Utilized fluorescence microscopy for LD imaging and analysis.
Main Results:
- Confirmed that BODIPY 493/503 can exhibit red fluorescence, potentially confounding LD analysis.
- The proposed protocol effectively prevents the dye from emitting erroneous red fluorescence.
- Ensured accurate green fluorescence signal for reliable LD visualization.
Conclusions:
- The spectral variability of BODIPY 493/503 necessitates careful experimental design.
- The developed protocol enhances the reliability of LD fluorescence labeling.
- Accurate lipid droplet imaging is critical for advancing cell biology research.
