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

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.

Auto-Phase-Locked Time-Resolved Luminescence Detection: Principles, Applications, and Prospects

“…In addition to the excitation sources and detectors, precise circuit systems with small timing jitter are usually used to synchronize the camera or shutter with the pulse excitation in many time-resolved detection systems (Krishnan et al, 2003;Urayama et al, 2003;Biskup et al, 2004;Connally et al, 2006;Qu et al, 2006;Sun et al, 2009;Gahlaut and Miller, 2010;Cicchi and Pavone, 2011;Strat et al, 2011;Becker, 2012;Hirvonen et al, 2014;Bergmann et al, 2016;Pominova et al, 2016;Chen T. et al, 2017;Luo et al, 2017;Liu et al, 2019Liu et al, , 2020. For Damayanti et al, 2013;Lu et al, 2014;Wang et al, 2017 Streak camera Scanning <1 ps 0.26 ns−1 ms Krishnan et al, 2003;Biskup et al, 2004;Qu et al, 2006;Bergmann et al, 2016;Pominova et al, 2016 Intensified camera Wide-field 0.2 ns 0.6 ns−1 ms Urayama et al, 2003;Connally et al, 2006;Sun et al, 2009;Gahlaut and Miller, 2010;Hirvonen et al, 2014;Chen T. et al, 2017;Liu et al, 2020 Gated detecting the delayed luminescence within nanoseconds, the related instruments are always composed of elements, such as ultrafast lasers, high-speed cameras and precise control circuits, which are very complicated and expensive to be popularized. However, there is no need to detect the microsecond-delay luminescence with nanosecond shutters, and a wide range of technologies were developed in the past decades to reduce the cost for time-resolved detection of phosphorescence and delayed fluorescence (Marriott et al, 1991(Marriott et al, , 1994...…”

“…A time-resolved luminescence detection instrument is usually composed of a pulse source, an optical detector and a synchronous control component (Krishnan et al, 2003 ; Urayama et al, 2003 ; Biskup et al, 2004 ; Connally et al, 2006 ; Qu et al, 2006 ; Connally and Piper, 2008 ; Sun et al, 2009 ; Gahlaut and Miller, 2010 ; Cicchi and Pavone, 2011 ; Strat et al, 2011 ; Becker, 2012 ; Damayanti et al, 2013 ; Grichine et al, 2014 ; Hirvonen et al, 2014 ; Lemmetyinen et al, 2014 ; Lu et al, 2014 ; Bergmann et al, 2016 ; Pominova et al, 2016 ; Bui et al, 2017 ; Chen T. et al, 2017 ; Luo et al, 2017 ; Wang et al, 2017 ; Zhang et al, 2018 ; Zhu et al, 2018b ; Liu et al, 2019 , 2020 ). To achieve a temporal resolution of nanoseconds, many instruments equipped with a picosecond or even femtosecond laser (Krishnan et al, 2003 ; Urayama et al, 2003 ; Biskup et al, 2004 ; Qu et al, 2006 ; Sun et al, 2009 ; Grichine et al, 2014 ; Pominova et al, 2016 ; Bui et al, 2017 ; Luo et al, 2017 ; Zhu et al, 2018b ), which is expensive. In order to increase the repetition rates and shorten the acquisition time, some lasers usually reach a frequency up to dozens of MHz (Damayanti et al, 2013 ; Luo et al, 2017 ), which is however not suitable for detecting luminescence with microseconds delay.…”

“…To achieve the time-resolved luminescence detection, the temporal resolution of related instruments should be high enough, since most luminescence probes exhibit luminescence lifetimes rang from sub-nanoseconds to milliseconds (Berezin and Achilefu, 2010 ; Bünzli, 2010 ; Yang et al, 2013 ; Zhang et al, 2018 ). With the development of pulse light sources, optical detectors and high-speed shutters, many instruments could achieve nanosecond-resolved luminescence detection (Krishnan et al, 2003 ; Urayama et al, 2003 ; Biskup et al, 2004 ; Connally et al, 2006 ; Qu et al, 2006 ; Sun et al, 2009 ; Gahlaut and Miller, 2010 ; Strat et al, 2011 ; Damayanti et al, 2013 ; Grichine et al, 2014 ; Hirvonen et al, 2014 ; Lu et al, 2014 ; Bergmann et al, 2016 ; Pominova et al, 2016 ; Bui et al, 2017 ; Luo et al, 2017 ; Wang et al, 2017 ; Zhu et al, 2018b ; Liu et al, 2019 ). Photon sensitivity is another important factor in time-resolved luminescence detections.…”

“…Because of these requirements, high-speed and sensitive detectors, such as PMT (photomultiplier tube) (Strat et al, 2011 ; Grichine et al, 2014 ; Bui et al, 2017 ; Luo et al, 2017 ; Zhu et al, 2018b ), SPAD (single-photon avalanche diode) (Damayanti et al, 2013 ; Lu et al, 2014 ; Wang et al, 2017 ), streak camera (Krishnan et al, 2003 ; Biskup et al, 2004 ; Qu et al, 2006 ; Bergmann et al, 2016 ; Pominova et al, 2016 ) or intensified camera (Urayama et al, 2003 ; Connally et al, 2006 ; Sun et al, 2009 ; Gahlaut and Miller, 2010 ; Hirvonen et al, 2014 ) are essential in many instruments for time-correlated single-photon counting (TCSPC). In addition, theses detectors always need to be synchronized with the ultrafast laser sources, leading to highly precise and complicated optical systems.…”

“…Most of these techniques are mainly used in detecting nanosecond-delayed fluorescence. As the developments of phosphorescence, delayed fluorescence and upconversion luminescence materials, these materials could emit luminescence with a delay time over microseconds or milliseconds (Marriott et al, 1991 , 1994 ; Verwoerd et al, 1994 ; Vereb et al, 1998 ; Connally et al, 2006 ; Connally and Piper, 2008 ; Bünzli, 2010 ; Gahlaut and Miller, 2010 ; Connally, 2011 ; Jin, 2011 ; Jin and Piper, 2011 ; Damayanti et al, 2013 ; Yang et al, 2013 , 2019 ; Grichine et al, 2014 ; Hirvonen et al, 2014 ; Jin et al, 2014 ; Lu et al, 2014 ; Zhang et al, 2014 , 2018 ; Bergmann et al, 2016 ; Pominova et al, 2016 ; Zheng et al, 2016 ; Zhu et al, 2016 , 2018a , b ; Bui et al, 2017 ; Chen T. et al, 2017 ; Wang et al, 2017 ; Sakiyama et al, 2018 ; Zhu and Shu, 2018 , 2019 ; Deng et al, 2020 ; Liu et al, 2020 ), greatly reducing the requirement of temporal resolution and the cost of the instruments. Combining with the use of low-cost shutters and automatic synchronization methods, CW light sources and common cameras were successfully used to accomplish the time-resolved luminescence detection with the temporal resolution ranging from microseconds to milliseconds (Marriott et al, 1991 , 1994 ; Verwoerd et al, 1994 ; Vereb et al, 1998 ; Connally, 2011 ; Jin, 2011 ; Jin and Piper, 2011 ; Jin et al, 2014 ; Zhang et al, 2014 ; Zheng et al, 2016 ; Sakiyama et al, 2018 ; Zhu and Shu, 2018 , 2019 ; Zhu et al, 2018b ; Yang et al, 2019 ; Deng et al, 2020 ).…”