3D sub-cellular localization of upconverting nanoparticles through hyperspectral microscopy


Hyperspectral microscopy is an intriguing technique combining spectroscopy with optical microscopy that can be used to simultaneously obtain spectral and spatial information. The relevance of hyperspectral imaging in biomedical applications such as the monitoring of bioimaging agents, the identification of pathogens and cancerous cells, and the cellular uptake of nanoparticles has emerged recently, due to recent advances in optical reconstruction. The location and tracking of particles within the cell structure have been analyzed by 2D hyperspectral imaging of non-fluorescence objects, being examples of 3D localization uncommon. Here, we report the synthesis of Yb3+/Er3+-codoped Gd2O3 nanoparticles, their structural and luminescence characterization, and their biocompatibility assessments in Human melanoma (MNT-1 and A375) cell lines. The internalization of the particles by MNT-1 cells and their 3D localization in a fixed configuration are addressed through 2D optical images acquired in different planes along with the cell culture depth. 2D hyperspectral imaging is used to unequivocally identify the nuclei and the nanoparticles. The results indicate that the particles are distributed in distinct planes deep in the cell volume in the cytoplasmic and perinuclear regions. Furthermore, the emission signature of the nanoparticles enabled the determination of the intracellular temperature.






Silva, RN; Botas, AMP; Brandao, D; Bastos, V; Oliveira, H; Debasu, ML; Ferreira, RAS; Brites, CDS; Carlos, LD

nossos autores


The work was partially developed under the project CICECO - Aveiro Institute of Materials, UIDB/50011/2020 & UIDP/50011/2020, and CESAM, UIDP/50017/2020 & UIDB/50017/2020, UID/QUI/00062/2019, financed by Portuguese funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement. Financial support from the projects NanoTBTech (European Union's Horizon 2020 FET Open program under grant agreement no. 801305) , NanoHeatControl (POCI-01-0145-FEDER-031469) , and HOT-sPOT (POCI-01-0145-FEDER-031794) , the last both funded by FEDER and Portuguese funds through FCT/MCTES, are acknowledged. RNS and AMPB thank NanoHeatControl for the grants.

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