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Mapping Molecular Diffusion in the Plasma Membrane by Multiple-Target Tracing MTT

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Published on: May 27, 2012

Simultaneous multi-species tracking in live cells with quantum dot conjugates.

Mathias P Clausen¹, Eva C Arnspang¹, Byron Ballou²

¹MEMPHYS - Center for Biomembrane Physics and Danish Molecular Biomedical Imaging Center (DaMBIC), University of Southern Denmark, Odense M, Denmark.

|June 4, 2014

Summary

Quantum dots enabled simultaneous tracking of membrane proteins and lipids. Their distinct diffusion patterns reveal cell membrane heterogeneity, which is reduced by methyl-β-cyclodextrin treatment.

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Area of Science:

Biophysics
Cell Biology
Nanotechnology

Background:

Quantum dots offer tunable optical properties and surface functionalization for biological imaging.
Investigating the lateral dynamics of multiple membrane components simultaneously is crucial for understanding cell function.
Previous studies have analyzed individual membrane protein or lipid dynamics, but not their concurrent behavior.

Purpose of the Study:

To simultaneously investigate the lateral dynamics of transmembrane epidermal growth factor receptor, CD59, and ganglioside GM1-cholera toxin subunit B clusters in the plasma membrane.
To determine if single trajectory analysis is robust for membrane dynamics studies.
To explore the effect of methyl-β-cyclodextrin on membrane component diffusion.

Main Methods:

Utilizing spectrally distinct, water-stabilized quantum dots for multiplexed single-particle tracking.
Performing simultaneous live-cell imaging of quantum dot-labeled biomolecules.
Analyzing single particle trajectories to extract diffusion coefficients and characterize heterogeneity.
Conducting simulations to validate the sufficiency of trajectory length for analysis.
Treating cells with methyl-β-cyclodextrin to observe its effects on membrane dynamics.

Main Results:

Demonstrated the feasibility of simultaneous tracking of three distinct plasma membrane species.
Confirmed that trajectories longer than 50 steps are sufficient for robust diffusion analysis.
Revealed heterogeneous distributions of diffusion coefficients for all three species, with species-specific differences.
Observed a decrease in diffusion heterogeneity upon methyl-β-cyclodextrin treatment.

Conclusions:

Quantum dots provide a versatile platform for multi-component membrane dynamics studies.
Plasma membrane organization exhibits distinct diffusion characteristics for different molecular species.
Cholesterol depletion using methyl-β-cyclodextrin alters membrane fluidity and reduces diffusion heterogeneity.