A small molecule two-photon probe for hydrogen sulfide in live tissues

We report a two-photon probe (FS1) which shows a 21-fold two-photon excited fluorescence enhancement in response to H(2)S and can selectively detect H(2)S in a rat hippocampal slice at a depth of 90-190 μm by using two-photon microscopy.     

Organically modified silica nanoparticles co-encapsulating photosensitizing drug and aggregation-enhanced two-photon absorbing fluorescent dye aggregates for two-photon photodynamic therapy

We report energy-transferring organically modified silica nanoparticles for two-photon photodynamic therapy. These nanoparticles co-encapsulate two-photon fluorescent dye nanoaggregates as an energy up-converting donor and a photosensitizing PDT drug as an acceptor. They combine two features: (i) aggregation-enhanced two-photon absorption and emission properties of a novel two-photon dye and (ii) nanoscopic fluorescence resonance energy transfer between this nanoaggregate and a photosensitizer, 2-devinyl-2-(1-hexyloxyethyl)pyropheophorbide. Stable aqueous dispersions of the co-encapsulating nanoparticles (diameter < or = 30 nm) have been prepared in the nonpolar interior of micelles by coprecipitating an organically modified silica sol with the photosensitizer and an excess amount of the two-photon dye which forms fluorescent aggregates by phase separation from the particle matrix. Using a multidisciplinary nanophotonic approach, we show: (i) indirect excitation of the photosensitizer through efficient two-photon excited intraparticle energy transfer from the dye aggregates in the intracellular environment of tumor cells and (ii) generation of singlet oxygen and in vitro cytotoxic effect in tumor cells by photosensitization under two-photon irradiation.     

A highly sensitive two-photon fluorescent probe for mitochondrial zinc ions in living tissue

We report a highly sensitive two-photon probe (SZn2-Mito) which shows a 70-fold two-photon excited fluorescence enhancement in response to Zn(2+) and can selectively detect mitochondrial Zn(2+) in a rat hippocampal slice at a depth of 100-200 μm by using two-photon microscopy.     

Two-photon fluorescent probe for cadmium imaging in cells

A novel two-photon excited fluorescent probe for cadmium (named as TPCd) was designed and synthesized utilizing a prodan (6-acetyl-2-methoxynaphthalene) derivative as the two-photon fluorophore and an o-phenylenediamine derivative as the Cd(2+) chelator, which possessed favorable photophysical properties and good water-solubility. The probe was designed with a photoinduced electron transfer (PET) mechanism and thus was weakly fluorescent itself. After binding with Cd(2+) which blocked the PET process, the fluorescence intensity of the probe was enhanced by up to 15-fold under one-photon excitation (OPE) and 27-fold under two-photon excitation (TPE), respectively. The two-photon action cross-section (Φδ) of the TPCd-Cd complex at 740 nm reached 109 GM compared to 3.6 GM for free TPCd, indicating the promising prospect of the probe in two-photon application. TPCd chelated Cd(2+) with 1 : 1 stoichiometry, and the apparent dissociation constant (K(d)) was 6.1 × 10(-5) M for the one-photon mode and 7.2 × 10(-5) M for the two-photon mode. The probe responded to Cd(2+) over a wide linear range from 0.1 to 30 μM with a detection limit of 0.04 μM. High selectivity of the probe towards Cd(2+) was acquired in Tris-HCl/sodium phosphate buffer. The probe was pH-independent in the biologically relevant pH range and non-toxic to living cells at reasonable concentration levels, warranting its in vivo applications. Through two-photon microscopy imaging, the probe was successfully applied to detect Cd(2+) uptake in living HepG2 cells.     

双氰基二苯代乙烯型双光子荧光汞离子探针

分别以双氰基二苯代乙烯(DCS)和双[2-(2-羟乙基硫基)乙基]氨(HSA)为双光子荧光团和汞离子受体,合成了双光子荧光汞离子探针(DHg),并对其结构进行了分析.实验结果表明,DHg在甲苯、乙腈和水中的荧光量子产率(Φ)分别为0.78,0.42和0.20,对汞离子的络合常数通过单、双光子荧光滴定分别拟合为lg K=5.47±0.02和lg K=5.34±0.02.DHg在水溶液中对汞离子具有优良的选择性和高的灵敏性,可用于中性环境中汞离子的检测.DHg的双光子吸收截面(δTPA)在水溶液中高达840 GM,可用于细胞中汞离子的检测与成像.     

Two-photon fluorescent probes for long-term imaging of calcium waves in live tissue

2-Acetyl-6-(dimethylamino)naphthalene-derived two-photon fluorescent Ca2+ probes (ACa1-ACa3) are reported. They can be excited by a 780 nm laser beam, show 23-50-fold enhancement in one- and two-photon excited fluorescence in response to Ca2+, emit fourfold stronger two-photon excited fluorescence than Oregon Green 488 BAPTA-1 upon complexation with Ca2+, and can selectively detect intracellular free Ca2+ ions in live cells and living tissues with minimum interference from other metal ions and membrane-bound probes. Moreover, these probes are capable of monitoring calcium waves at a depth of 120-170 microm in live tissues for 1100-4000 s using two-photon microscopy with no artifacts of photobleaching.     

Molecular two-photon sensor for metal ions derived from bis(2-pyridyl)amine

A molecular two-photon sensor for the metal ions derived from bis(2-pyridyl)amine as the receptor is reported. The sensor emits strong two-photon fluorescence when excited by 780 nm laser photons. Moreover, the binding constants measured by the one- and two-photon fluorescence are similar. This result may be useful for the design of efficient two-photon fluorescence probe for biological substrates.     

Dissociation of sulfur dioxide by ultraviolet multiphoton absorption between 224 and 232 nm

Multiphoton excitation and dissociation of SO(2) have been investigated in the wavelength range from 224 to 232 nm. Strong evidence is found for two-photon excitation to the H Rydberg state, followed by dissociation to SO + O and ionization of the SO product by absorption of a third photon. The two-photon excitation is resonantly enhanced via the C (1)B(2) intermediate state, and the two-photon yield spectrum thus bears a strong resemblance to the spectrum of this intermediate. Imaging of the O((3)P(2)), S((1)D(2)), and SO products suggests that, following dissociation of SO(2) from the H state, SO is produced in the A and B electronic states. S((1)D(2)) is produced both from two-photon dissociation of SO(2) to give S((1)D(2)) + O(2) and by single-photon dissociation of SO(+). In the former process, the O(2) is likely formed in all of its lowest three electronic states.     

1,2,3,4-四氢喹啉-4-酮衍生物的线性及非线性光学性质

合成了一系列具有刚性结构的推拉型1,2,3,4-四氢喹啉-4-酮衍生物: 1-苄基-1,2,3,4-四氢喹啉-4-酮(BTHQ)、2-(1,2,3,4-四氢喹啉-4-叶立德)丙二腈(THQM)、1,6-二羰基久洛尼定(DOJ)和1,6-二(二氰甲烯基叶立德)久洛尼定(BDCJ).测定了其吸收光谱、单光子荧光光谱和双光子上转换荧光光谱. 这类化合物的单双光子荧光参数都存在着很强的、规则的溶剂效应, 表明分子激发态可能存在较大的极性. 它们的二氯甲烷溶液在800 nm飞秒激光(150 fs)照射下, 能够发射出很强的双光子上转换荧光. 采用非线性透过率法测得四个化合物的双光子吸收截面在0.83～23.95×10－50 cm4&#8226;s&#8226;photon－1之间. 这类化合物的激发态存在有效的分子内电荷转移, 对双光子吸收和双光子荧光发射有较大贡献.     

Impressive europium red emission induced by two-photon excitation for biological applications

Three triazine-based europium(III) complexes were synthesized that demonstrated strong two-photon induced europium emission with a high two-photon absorption cross-section. The modified triazine ligand of complex 3 initiated over 100% enhancement of the two-photon absorption cross-section (σ(2): 320 GM) when compared with complex 1 (σ(2): 128 GM) in a solution of DMSO. Europium complex 3 is also stable in vitro, and power-dependence curves were obtained in vitro to confirm the two-photon-induced f-f emission in HeLa cells.     

A novel quinoline-based two-photon fluorescent probe for detecting Cd2+ in vitro and in vivo

A new two-photon fluorescent Cd(2+) probe APQ is developed by introducing a N(1),N(1)-dimethyl-N(2)-(pyridin-2-ylmethyl)ethane-1,2-diamine binding group and a 4-methoxyphenylvinyl conjugation-enhancing group to the 2- and 6-positions of quinoline. This probe shows a large red shift and good emission enhancement under Cd(2+) binding. It also exhibits a high ion selectivity for Cd(2+) (especially over Zn(2+)) and a large two-photon absorption cross section at 710 nm. Two-photon microscopy imaging studies reveal that the new probe is non-toxic and cell-permeable and can be used to detect intracellular Cd(2+) under two-photon excitation.     

对位氨基能显著提高D-A-D型化合物的双光子吸收性能

报道了具有典型D-A-D型共轭结构的反式2,5-二氰基-1,4-二(4'-甲氧基苯乙烯基)苯(MOS-CN), 2,5-二氰基-1,4-二(4'-二甲胺基苯乙烯基)苯(MAS-CN)和1,4-二(4'-甲氧基苯乙烯基)苯(MOS)的合成. 用核磁、红外和元素分析进行了表征. 测试了紫外吸收光谱、单光子荧光光谱、双光子荧光光谱、双光子吸收系数及双光子吸收截面. 在800 nm的飞秒脉冲激光激发下, 化合物MOS-CN, MAS-CN和MOS分别发出很强的绿色、黄色和蓝色上转换荧光. 化合物MOS-CN, MAS-CN和MOS的最大吸收波长、单光子发射波长、双光子诱导荧光波长、荧光量子产率、双光子吸收系数、双光子吸收截面及双光子荧光寿命各分别是393, 473, 367 nm; 470, 569, 434 nm; 475, 574, 438 nm; 0.12, 0.72, 0.21; 0.8, 5.3, 0.3 cm/GW; 270, 1790, 101 GM; 140 ps, 1.32 ns, 54 ps. MAS-CN的双光子吸收截面是MOS-CN的6.63倍, MOS-CN的双光子吸收截面是MOS的2.67倍, 表明对位氨基显著地提高了化合物的双光子吸收性能, 氰基也较大地提高了双光子吸收截面.     

A two-photon fluorescent probe for ratiometric imaging of hydrogen peroxide in live tissue

We report a two-photon fluorescent probe (PN1) that can be excited by 750 nm femto-second pulses, shows high photostability and negligible toxicity, and can visualize H(2)O(2) distribution in live cells and tissue by two-photon microscopy.     

Synthesis, Characterization and Non-linear Optical Properties of a New Organic Dye： DEAHAS

Two-photon absorption (TPA) is a process in which two photons are simultaneously absorbed to an excited state via a virtual state. The synthesis of organic optical materials with large TPA cross section has become a subject of great interest in recent years due to various application such as three-dimensional fluorescence imaging1, optical data storage2,3 and lithographic microfabrication4-6. Recently we have synthesized a new organic dye DEAHAS that is a symmetrically substituted stilbe…     

Metal ion sensing novel calix[4]crown fluoroionophore with a two-photon absorption property

1,3-Alternate calix[4]arene-based fluorescent chemosensors bearing two-photon absorbing chromophores have been synthesized, and their sensing behaviors toward metal ions were investigated via absorption band shifts as well as one- and two-photon fluorescence changes. Free ligands absorb the light at 461 nm and weakly emit their fluorescence at 600 nm when excited by UV-vis radiation at 461 nm, but no two-photon excited fluorescence is emitted by excitation at 780 nm. Addition of an Al(3+) or Pb(2+) ion to a solution of the ligand causes a blue-shifted absorption and enhanced fluorescence due to a declined resonance energy transfer (RET) upon excitation by one- and two-photon processes. Addition of a Pb(2+) ion to a solution of 1.K(+) results in a higher fluorescence intensity than the original 1.Pb(2+) complex regardless of one- or two-photon excitation, due to the allosteric effect induced by the complexation of K(+) with a crown loop.     

氧芴三苯胺多枝分子的双光子吸收与电化学行为

研究了3个氧芴/三苯胺衍生物: E-2,8-双(4-二苯胺基苯乙烯基)氧芴(简称OT-G1)、E-2,8-双[4-(二苯基氨基-二苯乙烯基)(4’-溴苯基)氨基-苯乙烯基]氧芴(简称OT-G1.5)和E-2,8-双-[4’,4″-二-(二苯胺基苯乙烯基)-4-二苯胺基苯乙烯基]氧芴(简称OT-G2)的双光子吸收和电化学行为. 研究结果表明, 分子“代数”从1→1.5→2增高, 氧芴三苯胺多枝分子的HOMO能级升高、双光子荧光强度和双光子吸收截面明显增大. 由于HOMO能级的升高有利于分子的电荷转移, 因而分子表现出强的双光子吸收能力, 这表明可通过电化学行为来推断出分子的双光子吸收性能.     

Two-photon fluorescent turn-on probe for lipid rafts in live cell and tissue

We report a new two-photon fluorescence turn-on probe 6-[(E)-3-oxo-1-dodecenyl]-2-[N-methyl-N-(carboxymethyl)amino]naphthalene (CL2) that is designed specifically for visualizing lipid rafts in living cells and tissues. This probe emits much brighter two-photon excited fluorescence in lipid rafts than in non-raft domains and allows direct visualization of the lipid rafts in the live cells and pyramidal neuron layer of the CA1 region at a depth of 100-250 mum in live tissues using two-photon microscopy.     

Magnesium ion selective two-photon fluorescent probe based on a benzo[h]chromene derivative for in vivo imaging

A novel, two-photon probe for the detection of free Mg2+ ions in living cells and live tissues has been developed. The probe can be excited by 880 nm laser photons, emits strong two-photon excited fluorescence in response to Mg2+ ions, can be easily loaded into the cell and tissue, shows high photostability, and can measure the Mg2+ ion concentration without interference by Ca2+ ions in living cells. The intracellular dissociation constant (Kdi) for Mg2+ determined by the two-photon process is 2.5 mM, which is suitable for dynamic Mg2+ concentration measurement. In addition, the probe is capable of imaging endogenous stores of free Mg2+ at a few hundred micrometers depth in live tissues using two-photon microscopy (TPM).     

香豆素肟酯类双光子引发剂的合成及性能研究

开发高效的双光子引发剂是提升双光子聚合速度的关键。本文基于光致脱羧机制,设计并合成了两种以共轭香豆素作为生色团、肟酯作为引发基团的双光子引发剂,并通过实验测试结合模拟计算对该类引发剂的光物理和光化学行为进行了研究。结果表明,该类引发剂在400~500 nm区域具有较强吸收,在LED可见光辐照下发生分解,具有光漂白特性,光解后释放的活性种可引发丙烯酸酯类单体聚合。利用双光子三维微纳成型技术,该类香豆素肟酯化合物可有效用于构建高分辨率的三维微纳结构。并通过量子化学计算,对该类引发剂的引发机理进行了探讨。     

双光子激发的超分子聚合物纳米颗粒生物成像研究

将具有双-2-脲基-4[1H]-嘧啶酮（bisUPy）的β-二羰基氟硼类衍生物（BF₂-bisUPy）及卟啉衍生物（Por（Pt）-bisUPy）通过四重氢键作用组装成超分子聚合物,通过微乳液法制备成在水中均匀分散的超分子聚合物纳米颗粒（SPNP）。扫描电子显微镜形貌表征表明获得的纳米颗粒粒径约为60 nm。紫外-可见吸收光谱、荧光发射光谱及寿命衰减实验均证明纳米颗粒内BF₂-bisUPy与Por（Pt）-bisUPy可发生高效的能量传递。具有双光子吸收的BF₂-bisUPy作为能量供体,可通过荧光共振能量传递（FRET）增强双光子激发下Por（Pt）-bisUPy的发光。双光子激发荧光强度与激光功率测试表明所制备的超分子聚合物纳米颗粒具有强烈的双光子激发下的荧光及磷光双发射,这种纳米材料可进入细胞,具有优秀的生物相容性,并在双光子激发时表现出强烈的荧光和磷光双发射生物成像。