Molecular Switches and Multiple Logic Gates Based on 4-(2-Pyridylazo)resorcinol

A simple‐structured 4‐(2‐pyridylazo)resorcinol (PAR) system presents interesting properties with dual fluorescent outputs. Modulated by solution pH two kinds of reversible switch behaviors, "ON‐OFF" and "OFF‐ON", were realized with the PAR system. Stimulated by different combination of external stimulus, such as metal ions, UV irradiation and solution pH, the PAR system could perform multiple logic functions including three inputs AND, two inputs INHIBIT and combinatorial "NOR/AND" in parallel. The operation of the designed system is very simple and detected with a high sensitive fluorescent signal.     

A fluorescent molecular sensor for pH windows in traditional and polymeric biocompatible micelles: comicellization of anionic species to shift and reshape the ON window

A new approach is presented to obtain fluorescent sensors for pH windows that work in water and under biomimetic conditions. A single molecule that features all-covalently linked components is used, thus making it capable of working as a fluorescent sensor with an OFF/ON/OFF response to pH value. The components are a tertiary amine, a pyridine, and a fluorophore (pyrene). The forms with both protonated bases or both neutral bases quench the pyrene fluorescence, whereas the form with the neutral pyridine and protonated amine groups is fluorescent. The molecular sensor is also equipped with a long alkyl chain to make it highly hydrophobic in all its protonated and unprotonated forms, that is, either when neutral or charged. Accordingly, it can be confined at any pH value either in traditional (i.e., low-molecular-weight) nonionic surfactant micelles or inside polymeric, biocompatible micellar containers. Relevant for future applications in vivo, thanks to its strong hydrophobicity, no leakage of the molecular sensor is observed from the polymeric biocompatible micelles. Due to the proximity of the pyridine and amine functions in the molecular structure and the poor hydration inside the micelles, the observed pK(a) values are low so that the ON window is positioned at very low pH values. However, the window can be shifted to biologically relevant values by comicellization of anionic species. In particular, in the micelles of the nonionic surfactant TritonX-100, a shift of the ON window to pH 4-6 is obtained by addition of the anionic sodium dodecyl sulphate surfactant, whose negative charge promotes the stability of the protonated forms of the pyridine and amine fragments. In the case of the polymeric micelles, we introduce the use of the amphiphilic polystyrene sulfonate anionic polyelectrolyte, the comicellization of which induces a shift and sharpening of the ON window that is centered at pH 4.     

Fluorescence Responses of the Protonation and Deprotonation Processes between Phenolate and Phenol within Rosamine

有 pyronine 酚骨骼的二基于 rosamine 的 pH 探针 1a 和 1b 被简单一步舞反应设计并且综合。pH 滴定实验向氢离子在 550-750 nm 附近在在离开上荧光回答附近显示出那探针 1a 和 1b 展览。pK _{探查 1a 的} 是 8.29,当因为氢,探查 1b 的增加到 12.1 时,在它内结合。选择、竞争的实验显示普通离子和氨基酸没介入他们有氢离子的排放。而且,共焦荧光灯成像证明探查 1a 能在 HeLa 和 Ges-1 房间作为线粒体 biomarker 被服务。     

A near-infrared fluorescent calcium probe: a new tool for intracellular multicolour Ca2+ imaging

We report a novel near-infrared fluorescent calcium probe (KFCA), which has good optical properties such as intense NIR fluorescence emission (670 nm, QY: 0.24), excellent ON/OFF ratio (120-fold), and good wavelength-compatibility with visible-light-emissive fluorophores (Fluo-4, DsRed2), and which is applicable for real-time dual-colour intracellular Ca(2+) imaging.     

Giant Amplification of Photoswitching by a Few Photons in Fluorescent Photochromic Organic Nanoparticles

Controlling or switching the optical signal from a large collection of molecules with the minimum of photons represents an extremely attractive concept. Promising fundamental and practical applications may be derived from such a photon‐saving principle. With this aim in mind, we have prepared fluorescent photochromic organic nanoparticles (NPs), showing bright red emission, complete ON–OFF contrast with full reversibility, and excellent fatigue resistance. Most interestingly, upon successive UV and visible light irradiation, the NPs exhibit a complete fluorescence quenching and recovery at very low photochromic conversion levels (<5 %), leading to the fluorescence photoswitching of 420±20 molecules for only one converted photochromic molecule. This “giant amplification of fluorescence photoswitching” originates from efficient intermolecular energy‐transfer processes within the NPs.     

Zinc(II) and PPi selective fluorescence OFF-ON-OFF functionality of a chemosensor in physiological conditions

A fluorescent chemosensor based on a quinoline derivative, L(2) (OFF state), selectively senses Zn(2+) by effective chelate-enhanced fluorescence (ON state), which further shows selectivity toward PPi over competing anions like Pi, AMP, and ATP via fluorescence quenching (OFF state) in a 100% aqueous HEPES buffer (pH 7.4). A plausible mode for the selective binding of PPi to 1 has been demonstrated by quantum mechanical density functional theory calculations and high-resolution mass spectrometry analysis.     

Strong Emission Enhancement in pH-Responsive 2:2 Cucurbit[8]uril Complexes

Organic fluorophores, particularly stimuli-responsive molecules, are very interesting for biological and material sciences applications, but frequently limited by aggregation- and rotation-caused photoluminescence quenching. A series of easily accessible bipyridinium fluorophores, whose emission is quenched by a twisted intramolecular charge-transfer (TICT) mechanism, is reported. Encapsulation in a cucurbit[7]uril host gave a 1:1 complex exhibiting a moderate emission increase due to destabilization of the TICT state inside the apolar cucurbituril cavity. A much stronger fluorescence enhancement is observed in 2:2 complexes with the larger cucurbit[8]uril, which is caused by additional conformational restriction of rotations around the aryl/aryl bonds. Because the cucurbituril complexes are pH switchable, this system represents an efficient supramolecular ON/OFF fluorescence switch.     

Molecular Logic Gates and Switches Based on 1,3,4‐Oxadiazoles Triggered by Metal Ions

Organic molecular devices for information processing applications are highly useful building blocks for constructing molecular‐level machines. The development of “intelligent” molecules capable of performing logic operations would enable molecular‐level devices and machines to be created. We designed a series of 2,5‐diaryl‐1,3,4‐oxadiazoles bearing a 2‐(para‐substituted)phenyl and a 5‐(o‐pyridyl) group (substituent X=NMe₂, OEt, Me, H, and Cl; 1 a – e ) that form a bidentate chelating environment for metal ions. These compounds showed fluorescence response profiles varying in both emission intensity and wavelength toward the tested metal ions Ni²⁺, Cu²⁺, Zn²⁺, Cd²⁺, Hg²⁺, and Pb²⁺ and the responses were dependent on the substituent X, with those of 1 d being the most substantial. The 1,3,4‐oxadiazole O or N atom and pyridine N atom were identified as metal‐chelating sites. The fluorescence responses of 1 d upon metal chelation were employed for developing truth tables for OR, NOR, INHIBIT, and EnNOR logic gates as well as “ON‐OFF‐ON” and “OFF‐ON‐OFF” fluorescent switches in a single 1,3,4‐oxadiazole molecular system.     

罗丹明基“OFF-ON”型荧光探针研究的一些新进展

综述了近年来罗丹明衍生物在分子探针方面研究的一些新进展, 系统阐述了该类探针分子的构筑及探针分子片段由内酰胺螺环构架(无荧光/OFF型)到开环蒽醌体系(有荧光/ON型)转化过程中, 伴随着荧光的变化在离子和小分子检测方面的应用.     

A reversible fluorescence “ON–OFF–ON” sensor for sequential detection of F− and Cu2+ ions and its application as a molecular-scale logic device and security keypad lock

A new azoimine receptor, R₁, was synthesized by Schiff base condensation of 4-(4-butylphenyl) azophenol and 2,6-diaminopyridine and acts as a colorimetric and fluorometric chemosensor for F^? and also toward Cu²⁺ ions in aqueous environment. UV–Vis absorption and fluorescent emission spectra were employed to study the sensing process. Emission study was performed to examine the dual sensing ability of the obtained probe with sequential addition of F^? followed by Cu²⁺ and vice versa. The receptor is an efficient “ON–OFF” fluorescent probe for the fluoride ion. Also, R₁ + F^? operated as an “OFF–ON” fluorescent sensor for Cu²⁺ ions. Considering emission intensity and absorption wavelength for F^? and Cu²⁺ ions, a molecular system was developed with the ability to mimic the functions of XNOR logic gating on the molecular level. In addition, R₁ behaved as a molecular security keypad lock with F^? and Cu²⁺ inputs. The keypad lock operation is particularly important, as the output of the system depends not only on the proper combination but also on the order of input signals, creating the correct password that can be used to “open” this molecular keypad lock through strong fluorescence emission at 460?nm.     

Real-time measurement of endosomal acidification by a novel genetically encoded biosensor

Genetically encoded fluorescent proteins are optimal reporters when used to monitor cellular processes as they can be targeted to any subcellular region by fusion to a protein of interest. Here, we present the pH-sensitive fluorescent protein E¹GFP which is ideally suited to monitor pH changes in dynamic intracellular structures in real time with high spatio temporal resolution. E¹GFP is a ratiometric pH indicator by emission with a pK close to 6.0. We describe an application of this novel pH reporter in the measurement of pH changes along the endo-lysosomal pathway. By fusing E¹GFP to the HIV-Tat protein which is endowed with cell-penetrating properties, we were able to monitor multi-step endocytosis from the initial cell-surface binding through to the intracellular endocytic network in real time. This represents a framework for the application of E¹GFP to the in situ detection of pH changes involved in dynamic biological phenomena. Figure The green fluorecent protein variant, E1GFP, is a ratiometric pH-indicator by emission with a pK close to 6.0 and is therefore particularly suitable for pH detection below neutrality. Upon excitation of the neutral state of the chromophore (~400-410 nm), E1GFP emission properties are strongly dependent on the environmental pH. We describe an application of this novel pH-reporter in the measurement of pH changes along the endo-lysosomal pathway. By fusing E1GFP to the HIV-Tat protein, which is endowed with cell-penetrating properties, we were able to monitor in real-time multi-step endocytosis from the initial cell-surface binding through to the intracellular endocytic network. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Different Steric‐Twist‐Induced Ternary Memory Characteristics in Nonconjugated Copolymers with Pendant Naphthalene and 1,8‐Naphthalimide Moieties

Herein, novel random copolymers PMNN and PMNB were designed and synthesized, and the memory devices Al/ PMNN and PMNB /ITO both exhibited ternary memory performance. The switching voltages of the OFF–ON1 and ON1–ON2 transitions for both memory devices are around −2.0 and −3.5 V, respectively, and the ON1/OFF, ON2/ON1 current ratios are both up to 10³. The observed tristable electrical conductivity switching could be attributed to field‐induced conformational ordering of the naphthalene rings in the side chains, and subsequent charge trapping by 1,8‐naphthalimide moieties. More interestingly, by adjusting the connection sites of 1,8‐naphthalimide moieties to tune the steric‐twist effect, different memory properties were achieved ( PMNN showed nonvolatile write once, read many (WORM) memory behavior, whereas PMNB showed volatile static RAM (SRAM) memory behavior). This result will offer a guideline for the design of different high‐performance multilevel memory devices by tuning the steric effects of the chemical moieties.     

Mechanical Switches of Fluorescence

Fluorescence switches are molecular systems containing a light-emitting fragment whose activity can be quenched/revived reversibly, at will,through an external parameter, i.e., a change of pH or the variation of the redox potential.Fluorescence switches can be static (the emission of the fluorophore isswitched ON/OFF by a bistable covalently linked control unit) or dynamic(the change in fluorescence is accompanied by an oriented molecular motion). Of the latterclass of switches, we will consider the cases (i) of a metal scorpionate and (ii) ofsystems in which a metal is reversibly translocated between two nonequivalent compartmentsof a ditopic ligand.     

新型有机荧光染料嵌合的核壳荧光纳米材料的研制

采用油包水的反相微乳液方法,首次以羊抗人免疫球蛋白(IgG)标记的异硫氰酸荧光素(FITC)为核材料,成功地制备了FITC的核壳荧光纳米颗粒,克服了采用传统方法制备核壳荧光纳米颗粒中存在的荧光染料泄露的问题.制备的这种核壳荧光纳米颗粒比细胞小很多,且具有生物亲和性,可为纳米生物传感器件提供新型材料.基于该核壳荧光纳米颗粒的标记方法也为生物医学提供了一种新型的非同位素分析方法.     

STED Imaging the Dynamics of Lysosomes by Dually Fluorogenic Si-Rhodamine

Super-resolution microscopy (SRM) imaging of the finite subcellular structures and subtle bioactivities inside organelles delivers abundant cellular information with high fidelity to unravel the intricate biological processes. An ideal fluorescent probe with precise control of fluorescence is critical in SRM technique like stimulated emission depletion (STED). Si-rhodamine was decorated with both targeting group and H⁺-receptor, affording the dually fluorogenic Si-rhodamine in which the NIR fluorescence was efficiently controlled by the coalescent of spirolactone-zwitterion equilibrium and PeT mechanism. The dually fluorogenic characters of the probe offer a perfect mutual enhancement in sensitivity, specificity and spatial resolution. Strong fluorescence only released in the existence of targeting protein at acidic lysosomal pH, ensured precisely tracking the dynamic of lysosomal structure and pH in living cells by STED.     

Chemical control of the DNA light switch: cycling the switch ON and OFF

The emission of the DNA light-switch complex [Ru(bpy)2(tpphz)]2+ (bpy = 2,2'-bipyridine, tpphz = tetrapyrido[3,2-a:2',3'-c:3' ',2' '-h:2' ',3' '-j]phenazine) can be reversibly turned ON and OFF over several cycles. The tpphz and taptp (taptp = 4,5,9,18-tetraazaphenanthreno[9,10-b] triphenylene) ligands in [Ru(bpy)2(tpphz)]2+ and [Ru(bpy)2(taptp)]2+, respectively, intercalate between the DNA bases, and a 50-fold increase in emission intensity of [Ru(bpy)2(tpphz)]2+ is observed upon DNA intercalation. The [Ru(bpy)2(tpphz)]2+ DNA light switch can be turned OFF statically in the presence of Co2+, Ni2+, and Zn2+, and the emission can be fully restored by the addition of EDTA. Cycling of the DNA light switch OFF and ON can be accomplished through the successive introduction of Co2+ and EDTA, respectively, to solutions of DNA-bound [Ru(bpy)2(tpphz)]2+. Owing to the absence of additional coordination sites, the emission of DNA-intercalated [Ru(bpy)2(taptp)]2+ is not quenched by transition metal ions in solution. To our knowledge, this work presents the first example of a reversible DNA light switch.     

An Activatable Near-Infrared Fluorescent Probe for Precise Detection of the Pulmonary Metastatic Tumors: A Traditional Molecule Having a Stunning Turn

An accurate detection of lung metastasis is of great significance for making better treatment choices and improving cancer prognosis, but remains a big challenge in clinical practice. In this study, we propose a reinventing strategy to develop a pH-activatable near-infrared (NIR) fluorescent nanoprobe, pulmonary metastasis tracer (denoted as PMT), based on assembly of NIR dye IR780 and calcium phosphate (CaP). By delicately tuning the intermolecular interactions during the assembly process and dye doping content, as well as the synthetic condition of probe, the fluorescence of PMT could be finely adjusted via the tumor acidity-triggered disassembly. Notably, the selected PMT9 could sharply convert subtle pH variations into a distinct fluorescence signal to generate high fluorescence ON/OFF contrast, dramatically reducing the background signals. Benefiting from such preferable features, PMT9 is able to precisely identify not only the tumor sites in orthotopic lung cancer models but also the pulmonary metastases in mice with remarkable signal-to-background ratio (SBR). This study provides a unique strategy to turn shortcomings of traditional dye IR780 during in vivo imaging into advantages and further expand the application of fluorescent probe to image lung associated tumor lesions.     

Fluorescent and Biocompatible Ruthenium-Coordinated Oligo(p-phenylenevinylene) Nanocatalysts for Transfer Hydrogenation in the Mitochondria of Living Cells

It is challenging to design metal catalysts for in situ transformation of endogenous biomolecules with good performance inside living cells. Herein, we report a multifunctional metal catalyst, ruthenium-coordinated oligo(p-phenylenevinylene) (OPV-Ru), for intracellular catalysis of transfer hydrogenation of nicotinamide adenine dinucleotide (NAD⁺) to its reduced format (NADH). Owing to its amphiphilic characteristic, OPV-Ru possesses good self-assembly capability in water to form nanoparticles through hydrophobic interaction and π–π stacking, and numerous positive charges on the surface of nanoparticles displayed a strong electrostatic interaction with negatively charged substrate molecules, creating a local microenvironment for enhancing the catalysis efficiency in comparison to dispersed catalytic center molecule (TOF value was enhanced by about 15 fold). OPV-Ru could selectively accumulate in the mitochondria of living cells. Benefiting from its inherent fluorescence, the dynamic distribution in cells and uptake behavior of OPV-Ru could be visualized under fluorescence microscopy. This work represents the first demonstration of a multifunctional organometallic complex catalyzing natural hydrogenation transformation in specific subcellular compartments of living cells with excellent performance, fluorescent imaging ability, specific mitochondria targeting and good chemoselectivity with high catalysis efficiency.     

Two-photon nano-PEBBLE sensors: subcellular pH measurements

Intracellular pH mapping is of great importance as it plays a critical role in many cellular events. Also, in tissue, pH mapping can be an indicator for the onset of cancer. Here we describe a biocompatible, targeted, ratiometric, fluorescent, pH sensing nano-PEBBLE (Photonic Explorer for Biomedical use with Biologically Localized Embedding) that is based on two-photon excitation. Two-photon excitation minimizes the photobleaching and cell autofluorescence drastically, leading to an increase in the signal-to-noise ratio. PEBBLE nanosensors provide a novel approach for introducing membrane impermeant dyes, like HPTS, into cells. We use both non-targeted and F3 peptide targeted PEBBLE nanosensors for intracellular pH measurement of 9L cells. The intracellular measurements suggest that the non-targeted nanosensors are mostly trapped in endosomes, whereas the F3 peptide targeting enables them to escape/avoid these acidic compartments. Combining the advantages of pH sensitive PEBBLE nanoparticles, including their specific targeting, with the advantages of two-photon microscopy provides an attractive and promising prospect for non-invasive real-time monitoring of pH inside cancer cells and tissues.     

A Metalloregulated Four‐State Nanoswitch Controls Two‐Step Sequential Catalysis in an Eleven‐Component System

The nanomechanical switch 1 with its three orthogonal binding motifs—the zinc(II) porphyrin, azaterpyridine, and shielded phenanthroline binding station—is quantitatively and reversibly toggled back and forth between four different switching states by means of addition and removal of appropriate metal‐ion inputs. Two of the four switching stages are able to initiate catalytic transformations (ON1, ON2), while the two others shut down any reaction (OFF1, OFF2). Thus, in a cyclic four‐state switching process the sequential transformation A + B + C → AB + C → ABC can be controlled, which proceeds stepwise along the switching states OFF1→ON1 (click reaction: A + B → AB )→OFF2→ON2 (Michael addition: AB + C → ABC )→OFF1. Two consecutive cycles of the sequential catalysis were realized without loss in activity in a reaction system with eleven different components.