SU-G-IeP4-08: Initial Investigations of Up-Converting Nanoparticles (UCNP) for 3D Tissue Imaging in Optical-ECT

Authors


Abstract

Purpose:

Near-IR absorptive up-converting nanoparticles (UCNPs) is a novel contrast for optical-ECT that allows auto-fluorescence-free 3D imaging of labeled cells in a matrix of large (∼1cm3) unsectioned normal tissue. This has the potential to image small metastases or dormant cells that is difficult with down-converting fluorescing dyes due to auto-fluorescence. The feasibility of imaging UCNP in agarose phantoms and a mouse lung is demonstrated, aided by a 3D-printed optical-ECT stage designed to excite UCNP in a mouse lung.

Methods:

The UCNP, NaYF4:Yb/Er (20/2%), studied in this work up-converts 980nm light to visible light peaking sharply at ∼540nm. To characterize the UCNP emission as a function of UCNP concentration, cylindrical 2.5%wt agarose phantoms infused with UCNP at concentrations of 25µg/mL and 50µg/mL were exposed to 1.5W 980nm laser coupled to an optical fiber. The fiber was held stably at 1cm above the stage via a custom 3D-printed stage. An optically cleared lung harvested from a BALBc mice was then injected with 100µL of 1mg/mL UCNP solution ex vivo. Tomographic imaging of the UCNP emission in lung was performed.

Results:

The laser beam tract is visualized within the agarose phantom. A line profile of UCNP emission at 25µg/mL versus 50µg/mL shows that increasing the UCNP concentration increases emission count. UCNPs injected into a cleared mouse lung disperse throughout the respiratory tract, allowing for visualization and 3D reconstruction. Excitation before and after UCNP injection shows the tissue exhibits no auto-fluorescence at 980nm, allowing clear view of the UCNP without any obscuring features such as conventional down-converting fluorescent tags.

Conclusion:

We confirm that up-conversion in tissue circumvents completely tissue auto-fluorescence, which allowed background-free 3D reconstruction of the UCNP distribution. We also confirm that raising the UCNP concentration increases emission and that UCNPs are retained in agarose samples during the optical clearing process.

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