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Biomolecular Imaging of Cellular Uptake of Nanoparticles using Multimodal Nonlinear Optical Microscopy

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Published on: May 16, 2022

Scanning two-photon microscopy with upconverting lanthanide nanoparticles via Richardson-Lucy deconvolution.

Christian F Gainer¹, Urs Utzinger, Marek Romanowski

¹The University of Arizona, Department of Biomedical Engineering, Tucson, Arizona, 85721-0240, USA.

Journal of Biomedical Optics

|August 17, 2012

Summary

We developed an image processing method to overcome blurring in fast-scanning microscopy caused by long nanoparticle luminescence decay. This technique enables clear imaging with upconverting lanthanide nanoparticles at low excitation power.

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Area of Science:

Biomedical Optics
Nanotechnology
Microscopy

Background:

Upconverting lanthanide nanoparticles offer advantages for bioimaging but suffer from long luminescence decay times.
This decay causes image blurring in fast-scanning microscopy, limiting their application in nondescanned imaging modes.

Purpose of the Study:

To develop and validate an image processing method to mitigate luminescence lifetime effects in upconverting nanoparticle microscopy.
To enable high-resolution, fast-scanning microscopy using functionalized nanoparticles at low excitation power.

Main Methods:

Application of Richardson-Lucy deconvolution to correct for luminescence decay in microscopy images.
Functionalization of nanoparticles using lipids for water dispersibility and conjugation to antibodies (e.g., epidermal growth factor receptor antibody).
Imaging experiments conducted at near-infrared excitation power densities significantly lower than traditional two-photon microscopy.

Main Results:

Achieved lateral resolution of ~1.2 μm and axial resolution of 5 μm or better.
Demonstrated successful imaging at scan rates comparable to traditional two-photon microscopy.
Utilized excitation power densities as low as 850 W/cm(2), orders of magnitude lower than typical two-photon imaging.

Conclusions:

Richardson-Lucy deconvolution effectively corrects for luminescence lifetime blurring in upconverting nanoparticle microscopy.
Lipid-functionalized nanoparticles are suitable for biological applications, enabling targeted imaging.
This approach facilitates high-resolution 3D functional tissue imaging with unprecedentedly low excitation power densities.