Upconversion microparticles as time-resolved luminescent probes for multiphoton microscopy: desired signal extraction from the streaking effect.
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.
A. M. Prokhorov General Physics Institute, Russian Academy of Sciences, Vavilov Street, 38, Moscow 119991, Russia.
Daria V
DV
Pominova
A. M. Prokhorov General Physics Institute, Russian Academy of Sciences, Vavilov Street, 38, Moscow 119991, RussiabNational Research Nuclear University MEPhI (Moscow Engineering Physics Institute), Kashirskoe Highway, 31, Moscow, 115409, Russia.
Anastasia V
AV
Ryabova
A. M. Prokhorov General Physics Institute, Russian Academy of Sciences, Vavilov Street, 38, Moscow 119991, Russia.
Pavel V
PV
Grachev
A. M. Prokhorov General Physics Institute, Russian Academy of Sciences, Vavilov Street, 38, Moscow 119991, Russia.
Igor D
ID
Romanishkin
A. M. Prokhorov General Physics Institute, Russian Academy of Sciences, Vavilov Street, 38, Moscow 119991, Russia.
Sergei V
SV
Kuznetsov
A. M. Prokhorov General Physics Institute, Russian Academy of Sciences, Vavilov Street, 38, Moscow 119991, Russia.
Julia A
JA
Rozhnova
A. M. Prokhorov General Physics Institute, Russian Academy of Sciences, Vavilov Street, 38, Moscow 119991, Russia.
Daria S
DS
Yasyrkina
A. M. Prokhorov General Physics Institute, Russian Academy of Sciences, Vavilov Street, 38, Moscow 119991, Russia.
Pavel P
PP
Fedorov
A. M. Prokhorov General Physics Institute, Russian Academy of Sciences, Vavilov Street, 38, Moscow 119991, RussiabNational Research Nuclear University MEPhI (Moscow Engineering Physics Institute), Kashirskoe Highway, 31, Moscow, 115409, Russia.
Victor B
VB
Loschenov
D006801:Humans; D007700:Kinetics; D049408:Luminescent Agents / Q000737:chemistry*; D018613:Microscopy, Confocal / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry*; D012815:Signal Processing, Computer-Assisted; D000074084:THP-1 Cells
D049408:Luminescent Agents