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Subdiffusive molecular motion in nanochannels observed by fluorescence correlation spectroscopy.

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¹Interdisciplinary Research Centre on Biomaterials (CRIB), University Federico II, Piazzale Tecchio 80, 80125, Naples, Italy.

Analytical Chemistry

|January 6, 2010

Summary

Confinement in nanochannels significantly slows large biomolecule diffusion. Flexible molecules exhibit subdiffusive motion, crucial for designing bioanalytic nanodevices.

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Area of Science:

Biophysics
Nanotechnology
Physical Chemistry

Background:

Understanding biomolecule behavior in confined spaces is essential for nanodevice development.
Molecular confinement influences diffusion dynamics, impacting transport and interactions.
Fluorescence Correlation Spectroscopy (FCS) is a powerful tool for studying single-molecule dynamics.

Purpose of the Study:

To investigate the impact of confinement on biomolecule motion within nanometric glass channels.
To quantify diffusion coefficients and concentrations as a function of molecule size and channel height.
To explore subdiffusive motion and its relationship with molecular flexibility and surface interactions.

Main Methods:

Utilized Fluorescence Correlation Spectroscopy (FCS) to analyze biomolecule motion.
Employed a single-component diffusion model to determine diffusion time and concentration.
Varied molecule size (Poly(ethylene glycol) 20 kDa, Dextran 40 kDa, Rhodamine 6G) relative to nanochannel height (r(g)/h).

Main Results:

Large molecules (PEG, Dextran) showed a ~10-fold decrease in diffusion coefficients as channel height approached molecular diameter.
Small molecules (Rhodamine 6G) were largely unaffected by nanochannel confinement.
Flexible molecules exhibited subdiffusive motion, with mobility deviating towards a square root dependence on time at r(g)/h ~ 0.5.
A model was developed to explain diffusion time dependence due to molecular rearrangements and surface dynamics.

Conclusions:

Confinement significantly alters biomolecule diffusion, with effects dependent on molecular size and flexibility.
Subdiffusive motion is a key characteristic of flexible molecules in nanochannels.
Extracted single-molecule parameters (partitioning, diffusion time, desorption, binding) are valuable for engineering bioanalytic nanodevices.