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Two-Dimensional Fluorescence Difference Spectroscopy to Characterize Nanoparticles and their Interactions.

Miranda N Hurst¹, Robert K DeLong¹

¹Nanotechnology Innovation Center Kansas State, Department of Anatomy and Physiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, USA.

Scientific Reports

|September 15, 2016

Summary

Two dimensional fluorescence difference spectroscopy (2D FDS) identifies nanoparticles and their surface interactions. This novel technique analyzes spectral signatures to ensure nanoparticle quality after functionalization and biomolecule loading.

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Area of Science:

Nanotechnology
Spectroscopy
Materials Science

Background:

Characterizing nanoparticle surface properties and biomolecule interactions is crucial for quality assurance.
Existing methods may lack the sensitivity or specificity required for comprehensive analysis.

Purpose of the Study:

To evaluate two dimensional fluorescence difference spectroscopy (2D FDS) as a novel method for identifying nanoparticles.
To assess 2D FDS's capability in detecting nanoparticle surface functionalization and biomolecule loading.

Main Methods:

Utilized 2D FDS to generate spectral signatures of various metal oxide nanoparticles (NPs).
Investigated changes in spectral signatures upon surface functionalization with glycol chitosan, polyacrylic acid (PAA), or methoxy polyethylene glycol (mPEG).
Analyzed spectral shifts in ZnO NPs loaded with Torula Yeast RNA (TYRNA), polyinosinic: polycytidylic acid (pIC), or splice switching oligonucleotide (SSO).
Determined the relationship between Ras-Binding domain (RBD) protein concentration and fluorescent intensity.

Main Results:

Each metal oxide nanoparticle composition exhibited a unique spectral signature.
2D FDS detected surface property variations in ZnO NPs synthesized via different methods.
Functionalization with different biomolecules (glycol chitosan, PAA, mPEG) caused distinct spectral signature shifts.
Loading with TYRNA, pIC, or SSO resulted in specific emission shifts.
Fluorescent intensity decreased proportionally with increasing concentrations of loaded RBD protein.

Conclusions:

2D FDS is a sensitive technique for differentiating nanoparticles based on material composition.
The method effectively identifies changes in nanoparticle surface properties and biomolecule interactions.
2D FDS serves as a valuable quality assurance tool for nanoparticle characterization.