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Journal of Biomedical Optics

Widefield imaging of upconverting nanoparticles on epifluorescence microscopes adapted for laser illumination with top-hat profile
Author(s): Jiri Mrazek; Martina Pospisilova; Vit Svozil; Ondrej Cadek; Kristina Nesporova; Romana Sulakova; Martina Brandejsova; Jana Vranova; Vladimir Velebny
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Paper Abstract

We describe a modification of epifluorescence microscopes that allows quantitative widefield imaging of samples labeled by upconverting nanoparticles (UCNP). A top-hat illumination profile on the sample was achieved with a 980-nm laser diode by using tandem microlens arrays, a moving diffuser and a telescope, which adjusts the top-hat area to the field of view. Illumination homogeneity is a critical factor for imaging of UCNP since the intensity of their luminescence typically scales with the second power of the excitation intensity. Our illuminator is combined with the epifluorescence attachment of the microscope, allowing easy switching between observation of UCNP and traditional fluorescent dyes. Illumination profile homogeneity of about 98% was measured for objectives with magnification from 4× to 100×, and the top-hat profile was also obtained with phase contrast objectives. We demonstrate capability of the illuminator by evaluating in vitro uptake of UCNP encapsulated in oleyl-hyaluronan micelles into breast cancer cells. Micelles bearing the targeting peptide were about an order of magnitude more efficient than nontargeted micelles.

Paper Details

Date Published: 27 May 2016
PDF: 9 pages
J. Biomed. Opt. 21(5) 056007 doi: 10.1117/1.JBO.21.5.056007
Published in: Journal of Biomedical Optics Volume 21, Issue 5
Show Author Affiliations
Jiri Mrazek, Charles Univ. in Prague (Czech Republic)
Contipro Group S.R.O. (Czech Republic)
Martina Pospisilova, Contipro Group S.R.O. (Czech Republic)
Vit Svozil, Contipro Group S.R.O. (Czech Republic)
Ondrej Cadek, Contipro Group S.R.O. (Czech Republic)
Kristina Nesporova, Contipro Group S.R.O. (Czech Republic)
Romana Sulakova, Contipro Group S.R.O. (Czech Republic)
Martina Brandejsova, Contipro Group S.R.O. (Czech Republic)
Jana Vranova, Charles Univ. in Prague (Czech Republic)
Vladimir Velebny, Contipro Group S.R.O. (Czech Republic)

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