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Updated: Oct 4, 2025
Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
Published on: June 30, 2018
Label-Free Live-Cell Imaging of Internalized Microplastics and Cytoplasmic Organelles with Multicolor CARS Microscopy
Dae Sik Choi1,2, Sohee Lim3,4, Jin-Sung Park3
1Technology Human Resource Support for SMEs Center, Korea Institute of Industrial Technology (KITECH), Cheonan 31056, Republic of Korea.
Abstract:
As the bioaccumulation of microplastics (MPs) is considered as a potential health risk, many efforts have been made to understand the cellular dynamics and cytotoxicity of MPs. Here, we demonstrate that label-free multicolor coherent anti-Stokes Raman scattering (CARS) microscopy enables separate vibrational imaging of internalized MPs and lipid droplets (LDs) with indistinguishable shapes and sizes in live cells. By simultaneously obtaining polystyrene (PS)- and lipid-specific CARS images at two very different frequencies, 1000 and 2850 cm-1, respectively, we successfully identify the local distribution of ingested PS beads and native LDs in Caenorhabditis elegans. We further show that the movements of PS beads and LDs in live cells can be separately tracked in real time, which allows us to characterize their individual intracellular dynamics. We thus anticipate that our multicolor CARS imaging method could be of great use to investigate the cellular transport and cytotoxicity of MPs without additional efforts for pre-labeling to MPs.
Insights
Label-free multicolor coherent anti-Stokes Raman scattering (CARS) microscopy visualizes microplastics (MPs) and lipid droplets in live cells. This technique tracks their intracellular dynamics, aiding MP toxicity research without pre-labeling.
Area of Science:
- Cellular biology
- Microscopy techniques
- Toxicology
Background:
- Microplastic (MP) bioaccumulation poses potential health risks.
- Understanding MP cellular dynamics and cytotoxicity is crucial.
- Current methods often require pre-labeling of MPs for imaging.
Purpose of the Study:
- To demonstrate a label-free method for visualizing MPs and cellular structures.
- To investigate the intracellular dynamics and distribution of MPs in live cells.
- To assess the potential of multicolor CARS microscopy for MP toxicity studies.
Main Methods:
- Utilized label-free multicolor coherent anti-Stokes Raman scattering (CARS) microscopy.
- Acquired simultaneous vibrational images of internalized polystyrene (PS) beads and lipid droplets (LDs).
- Tracked the real-time movements and intracellular dynamics of PS beads and LDs in *Caenorhabditis elegans*.
Main Results:
- Successfully visualized and distinguished internalized MPs from native LDs based on their vibrational signatures.
- Identified the precise spatial distribution of ingested PS beads and LDs within live cells.
- Characterized the individual intracellular dynamics and movements of MPs and LDs in real time.
Conclusions:
- Multicolor CARS microscopy offers a label-free approach to study MP-cell interactions.
- The method enables simultaneous imaging and tracking of MPs and endogenous cellular components like LDs.
- This technique can significantly advance research into MP cellular transport and cytotoxicity without pre-labeling.
