Oxygenation of tissues plays an important role in the development and progression of tumor to treatment effects.Method of metalloporphyrines phosphorescence quenching by oxygen is one of the ways to measure dynamics o...Oxygenation of tissues plays an important role in the development and progression of tumor to treatment effects.Method of metalloporphyrines phosphorescence quenching by oxygen is one of the ways to measure dynamics of the oxygen concentration in the tissues by phosphorescence lifetime imaging of meso-tetra(sulfophenyl)tetrabenzoporphyrin Pd(Ⅱ)(TBP)using the time-correlated single photon counting(TCSPC)method.It has been shown that phosphorescence lifetime of the sensor in S37 tumor in vivo varied in the range of 130 to 290μs after both topical and intravenous administration of TBP.It indicates that oxygen level in tumors was lower compared to normal tissues where TBP phosphorescence has not been detected.Phosphorescence lifetimes of TBP increased in the solid tumor and in the muscle after photodynamic therapy of solid tumor that demonstrates oxygen consumption during treatment and possibly stopping the blood flow and hence the oxygen supply to the tissues.展开更多
Fluorescence lifetime imaging microscopy(FLIM)is increasingly used in biomedicine,material science,chemistry,and other related research fields,because of its advantages of high specificity and sensitivity in monitorin...Fluorescence lifetime imaging microscopy(FLIM)is increasingly used in biomedicine,material science,chemistry,and other related research fields,because of its advantages of high specificity and sensitivity in monitoring cellular microenvironments,studying interaction between proteins,metabolic state,screening drugs and analyzing their efficacy,characterizing novel materials,and diagnosing early cancers.Understandably,there is a large interest in obtaining FLIM data within an acquisition time as short as possible.Consequently,there is currently a technology that advances towards faster and faster FLIM recording.However,the maximum speed of a recording technique is only part of the problerm.The acquisition time of a FLIM image is a complex function of many factors.These include the photon rate that can be obtained from the sample,the amount of information a technique extracts from the decay functions,the fficiency at which it determines fluorescence decay parameters from the recorded photons,the demands for the accuracy of these parameters,the number of pixels,and the lateral and axial resolutions that are obtained in biological materials.Starting from a discussion of the parameters which determine the acquisition time,this review will describe existing and emerging FLIM techniques and data analysis algo-rithms,and analyze their performance and recording speed in biological and biomedical applications.展开更多
We demonstrate a photon counting laser ranging experiment with a four-channel single-photon detector(SPD). The multi-channel SPD improve the counting rate more than 4×10~7 cps, which makes possible for the distan...We demonstrate a photon counting laser ranging experiment with a four-channel single-photon detector(SPD). The multi-channel SPD improve the counting rate more than 4×10~7 cps, which makes possible for the distance measurement performed even in daylight. However, the time-correlated single-photon counting(TCSPC) technique cannot distill the signal easily while the fast moving targets are submersed in the strong background. We propose a dynamic TCSPC method for fast moving targets measurement by varying coincidence window in real time. In the experiment, we prove that targets with velocity of 5 km/s can be detected according to the method, while the echo rate is 20% with the background counts of more than 1.2×10~7 cps.展开更多
Significantly reduced tissue scattering of fluorescence signals in the second near-infrared(NIR-Ⅱ,1,000–1,700 nm)spectral region offers opportunities for large-depth in vivo bioimaging.Nowadays,most reported works c...Significantly reduced tissue scattering of fluorescence signals in the second near-infrared(NIR-Ⅱ,1,000–1,700 nm)spectral region offers opportunities for large-depth in vivo bioimaging.Nowadays,most reported works concerning NIR-II fluorescence in vivo bioimaging are realized by wide-field illumination and 2D-arrayed detection(e.g.,via InGaAs camera),which has high temporal resolution but limited spatial resolution due to out-of-focus signals.Combining NIR-II fluorescence imaging with confocal microscopy is a good approach to achieve high-spatial resolution visualization of biosamples even at deep tissues.In this presented work,a NIR-II fluorescence confocal microscopic system was setup.By using a kind of aggregation-induced emission(AIE)dots as NIR-II fluorescent probes,800 lm-deep 3D in vivo cerebrovascular imaging of a mouse was obtained,and the spatial resolution at 700 lm depth could reach 8.78 lm.Moreover,the time-correlated single photon counting(TCSPC)technique and femtosecond laser excitation were introduced into NIR-II fluorescence confocal microscopy,and in vivo confocal NIR-II fluorescence lifetime microscopic imaging(FLIM)of mouse cerebral vasculature was successfully realized.展开更多
文摘Oxygenation of tissues plays an important role in the development and progression of tumor to treatment effects.Method of metalloporphyrines phosphorescence quenching by oxygen is one of the ways to measure dynamics of the oxygen concentration in the tissues by phosphorescence lifetime imaging of meso-tetra(sulfophenyl)tetrabenzoporphyrin Pd(Ⅱ)(TBP)using the time-correlated single photon counting(TCSPC)method.It has been shown that phosphorescence lifetime of the sensor in S37 tumor in vivo varied in the range of 130 to 290μs after both topical and intravenous administration of TBP.It indicates that oxygen level in tumors was lower compared to normal tissues where TBP phosphorescence has not been detected.Phosphorescence lifetimes of TBP increased in the solid tumor and in the muscle after photodynamic therapy of solid tumor that demonstrates oxygen consumption during treatment and possibly stopping the blood flow and hence the oxygen supply to the tissues.
基金support from the National Key R&D Program of China(2017YFA0700500)National Natural Science Foundation of China(61775144/61525503/61620106016/61835009/81727804)+2 种基金(Key)Project of Department of Education of Guangdong Province(2015KGJHZ002/2016KCXTD007)Guangdong Natural Science Foundation(2014A030312008,2017A030310132,2018A030313362)Shenzhen Basic Research Project(JCYJ20170818144012025/JCYJ20170818141701667/JCYJ20170412105003520/JCYJ20150930104948169).
文摘Fluorescence lifetime imaging microscopy(FLIM)is increasingly used in biomedicine,material science,chemistry,and other related research fields,because of its advantages of high specificity and sensitivity in monitoring cellular microenvironments,studying interaction between proteins,metabolic state,screening drugs and analyzing their efficacy,characterizing novel materials,and diagnosing early cancers.Understandably,there is a large interest in obtaining FLIM data within an acquisition time as short as possible.Consequently,there is currently a technology that advances towards faster and faster FLIM recording.However,the maximum speed of a recording technique is only part of the problerm.The acquisition time of a FLIM image is a complex function of many factors.These include the photon rate that can be obtained from the sample,the amount of information a technique extracts from the decay functions,the fficiency at which it determines fluorescence decay parameters from the recorded photons,the demands for the accuracy of these parameters,the number of pixels,and the lateral and axial resolutions that are obtained in biological materials.Starting from a discussion of the parameters which determine the acquisition time,this review will describe existing and emerging FLIM techniques and data analysis algo-rithms,and analyze their performance and recording speed in biological and biomedical applications.
基金supported by the National Natural Science Foundation of China(No.11374105)
文摘We demonstrate a photon counting laser ranging experiment with a four-channel single-photon detector(SPD). The multi-channel SPD improve the counting rate more than 4×10~7 cps, which makes possible for the distance measurement performed even in daylight. However, the time-correlated single-photon counting(TCSPC) technique cannot distill the signal easily while the fast moving targets are submersed in the strong background. We propose a dynamic TCSPC method for fast moving targets measurement by varying coincidence window in real time. In the experiment, we prove that targets with velocity of 5 km/s can be detected according to the method, while the echo rate is 20% with the background counts of more than 1.2×10~7 cps.
基金supported by the National Natural Science Foundation of China(61735016)Zhejiang Provincial Natural Science Foundation of China(LR17F050001)
文摘Significantly reduced tissue scattering of fluorescence signals in the second near-infrared(NIR-Ⅱ,1,000–1,700 nm)spectral region offers opportunities for large-depth in vivo bioimaging.Nowadays,most reported works concerning NIR-II fluorescence in vivo bioimaging are realized by wide-field illumination and 2D-arrayed detection(e.g.,via InGaAs camera),which has high temporal resolution but limited spatial resolution due to out-of-focus signals.Combining NIR-II fluorescence imaging with confocal microscopy is a good approach to achieve high-spatial resolution visualization of biosamples even at deep tissues.In this presented work,a NIR-II fluorescence confocal microscopic system was setup.By using a kind of aggregation-induced emission(AIE)dots as NIR-II fluorescent probes,800 lm-deep 3D in vivo cerebrovascular imaging of a mouse was obtained,and the spatial resolution at 700 lm depth could reach 8.78 lm.Moreover,the time-correlated single photon counting(TCSPC)technique and femtosecond laser excitation were introduced into NIR-II fluorescence confocal microscopy,and in vivo confocal NIR-II fluorescence lifetime microscopic imaging(FLIM)of mouse cerebral vasculature was successfully realized.
