Upconversion microparticles as time-resolved luminescent probes for multiphoton microscopy: desired signal extraction from the streaking effect

Daria V. Pominova; Anastasia Ryabova; Pavel N. Grachev; Igor D. Romanishkin; Sergey V. Kuznetsov; JUlia A. Rozhnova; Daria S. Yasyrkina; Pavel P. Fedorov; Victor B. Loschenov

doi:10.1117/1.JBO.21.9.096002

8 September 2016 Upconversion microparticles as time-resolved luminescent probes for multiphoton microscopy: desired signal extraction from the streaking effect

Daria V. Pominova, Anastasia Ryabova, Pavel N. Grachev, Igor D. Romanishkin, Sergey V. Kuznetsov, JUlia A. Rozhnova, Daria S. Yasyrkina, Pavel P. Fedorov, Victor B. Loschenov

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Journal of Biomedical Optics, Vol. 21, Issue 9, 096002 (September 2016). https://doi.org/10.1117/1.JBO.21.9.096002

Abstract

The great interest in upconversion nanoparticles exists due to their high efficiency under multiphoton excitation. However, when these particles are used in scanning microscopy, the upconversion luminescence causes a streaking effect due to the long lifetime. This article describes a method of upconversion microparticle luminescence lifetime determination with help of modified Lucy–Richardson deconvolution of laser scanning microscope (LSM) image obtained under near-IR excitation using nondescanned detectors. Determination of the upconversion luminescence intensity and the decay time of separate microparticles was done by intensity profile along the image fast scan axis approximation. We studied upconversion submicroparticles based on fluoride hosts doped with Yb³⁺-Er³⁺ and Yb³⁺-Tm³⁺ rare earth ion pairs, and the characteristic decay times were 0.1 to 1.5 ms. We also compared the results of LSM measurements with the photon counting method results; the spread of values was about 13% and was associated with the approximation error. Data obtained from live cells showed the possibility of distinguishing the position of upconversion submicroparticles inside and outside the cells by the difference of their lifetime. The proposed technique allows using the upconversion microparticles without shells as probes for the presence of OH− ions and CO₂ molecules.

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Daria V. Pominova, Anastasia Ryabova, Pavel N. Grachev, Igor D. Romanishkin, Sergey V. Kuznetsov, JUlia A. Rozhnova, Daria S. Yasyrkina, Pavel P. Fedorov, and Victor B. Loschenov "Upconversion microparticles as time-resolved luminescent probes for multiphoton microscopy: desired signal extraction from the streaking effect," Journal of Biomedical Optics 21(9), 096002 (8 September 2016). https://doi.org/10.1117/1.JBO.21.9.096002

Published: 8 September 2016

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