Spontaneously Blinking Rhodamine Dyes for Single-Molecule Localization Microscopy

Single-molecule localization microscopy (SMLM) has found extensive applications in various fields of biology and chemistry. As a vital component of SMLM, fluorophores play an essential role in obtaining super-resolution fluorescence images. Recent research on spontaneously blinking fluorophores has greatly simplified the experimental setups and extended the imaging duration of SMLM. To support this crucial development, this review provides a comprehensive overview of the development of spontaneously blinking rhodamines from 2014 to 2023, as well as the key mechanistic aspects of intramolecular spirocyclization reactions. We hope that by offering insightful design guidelines, this review will contribute to accelerating the advancement of super-resolution imaging technologies.     

原子力显微镜-荧光显微镜联用技术在活细胞单分子检测中的应用

原子力显微镜(Atomic force microscopy,AFM)及荧光显微镜(Fluorescence microscopy,FM)是目前活细胞单分子分析检测中最常用的两种工具.结合两种显微镜的优势,发展高时空分辨、多功能的AFM-FM联用技术成为近年该领域的研究热点.本文简述了AFM单分子力谱和FM单分子荧光成像的原理,总结了AFM-FM联用系统在仪器研制方面的发展概况,并结合本课题组在应用AFM-FM联用技术研究细胞膜上配受体相互作用等方面的工作,介绍了其在活细胞单分子检测中的应用进展.     

When Weak Is Strong: A Plea for Low-Affinity Binders for Optical Microscopy

The exploitation of low-affinity molecular interactions in protein labeling is an emerging topic in optical microscopy. Such non-covalent and low-affinity interactions can be realized with various concepts from chemistry and for different molecule classes, and lead to a constant renewal of fluorescence signals at target sites. Further benefits are a versatile use across microscopy methods, in 3D, live and many-target applications. In recent years, several classes of low-affinity labels were developed and a variety of powerful applications demonstrated. Still, this research field is underdeveloped, while the potential is huge.     

Industrial Applications of Photochemical Syntheses

The principal industrial applications of photochemistry have so far been in the fields of free-radical chlorination, sulfochlorination, sulfoxidation, and nitrosation. In addition, however, photochemical reactions are being utilized on an increasing scale for the synthesis of vitamins, drugs, and fragrances. The present article surveys the various kinds of light-induced reactions exploited industrially, the equipment developed, and uses of the photochemical products. Furthermore, the problems encountered in designing a photochemical production plant are discussed for the example of photonitrosation of cyclohexane.     

Photophysical and Photochemical Processes in Micellar Systems

A fascinating feature inherent to aqueous surfactant solutions is the phenomenon of self-organization: above a certain critical concentration (the critical micelle concentration, CMC) detergent molecules associate spontaneously to build up structural entities of colloidal dimensions called micelles. The architecture of these agglomerates is such that the interior contains the hydrophobic alkyl chain of the amphiphile while the hydrophilic head groups are located at the surface and are in contact with bulk water. In the case of ionic micelles the interface is charged giving rise to an electrical double layer and a potential difference of up to several hundred millivolts between the micellar pseudophase and water. Thus micellar systems are microheterogeneous in character: the electrostatic potential and polarity prevailing in the interior of the aggregate differ from those of the bulk aqueous phase. A particularly attractive aspect of photochemical studies in micellar systems is the possibility of organizing the reactants at a molecular level: by comparison of the data in micelles with similar data in homogeneous solution one can learn about the molecular details of a given reaction and establish which conditions favor one pathway or another. In simple surfactant systems differences in rate and efficiency of a reaction will often be controlled by local electrostatic potentials and the compartmentalization of the reagents within the surfactant aggregates. Through the latter effect the statistics of probe distribution over the micelles becomes important in controlling fast photochemical events. Functional micelles are distinguished by the fact that the surfactant molecule contains a group which itself participates in the photoprocess. These units are unique in that self-assembly often introduces striking cooperative effects.     

Photochemical transformations of xanthohumol

A photoinduced procedure for the addition of simple alcohols to xanthohumol, the major prenylated flavonoid of hop plants, has been developed. The reaction is regioselective, simple, and occurs in one pot to afford two new β adducts of xanthohumol.     

Generation of Carbenes by Photochemical Cycloelimination

A wide variety of shelf stable substrates undergo photochemical cycloelimination under relatively mild conditions. Suitable substrates include cyclopropanes, oxiranes, aziridines, and diazirines, as well as 3 H-pyrazoles and 1,3-dioxolanes. Carbenes prepared from these substrates as well as certain cyclic carbonates, sulfites, and phosphoranes behave chemically in a manner similar to divalent carbon species produced from conventional diazo precursors, namely, they react with alkenes to give cyclopropanes and undergo insertion into C? H bonds of alkanes. In many cases the carbenes formed may also be detected by EPR and optical spectroscopic methods.     

Novel Photochemical Rearrangement of Styrylfurans

A conrotatory photocyclization , a novel [1,9] hydrogen shift, and a lateral ring opening are involved in the rearrangement of a series of 2-(4-alkylstyryl)furans 1 to 5-(3-alkylbutadienyl)benzo[b]furans 2 in dichloromethane. These novel photochemical rearrangements occur with good yields of isolated products.     

Photochemical Schiemann Reaction in Ionic Liquids

Photochemical Schiemann reactions of imidazole derivatives 1 and 4 were carried out in 1-butyl-3-methylimidazolium tetrafluoroborate ionic liquid [bmim][BF₄] as solvent. The effects of temperature, co-solvent and wavelength on the rate of the reaction and product yield were examined. The use of ionic liquid increases the yield of the photochemical fluorodediazoniation reaction of 2 at 0 °C. Careful temperature control is necessary to minimize the photodecomposition of the ionic liquid in order to increase the yield of product.     

Long-Term,Single-Molecule Imaging of Proteins in Live Cells with Photoregulated Fluxional Fluorophores

Single-molecule localization microscopy (SMLM) can reveal nanometric details of biological samples, but its high phototoxicity hampers long-term imaging in live specimens. A significant part of this phototoxicity stems from repeated irradiations that are necessary for controlled switching of fluorophores to maintain the sparse labeling of the sample. Lower phototoxicity can be obtained using fluorophores that blink spontaneously, but controlling the density of single-molecule emitters is challenging. We recently developed photoregulated fluxional fluorophores (PFFs) that combine the benefits of spontaneously blinking dyes with photocontrol of emitter density. These dyes, however, were limited to imaging acidic organelles in live cells. Herein, we report a systematic study of PFFs that culminates in probes that are functional at physiological pH and operate at longer wavelengths than their predecessors. Moreover, these probes are compatible with HaloTag labeling, thus enabling timelapse, single-molecule imaging of specific protein targets for exceptionally long times.     

The Photochemical Synthesis of Fine Chemicals with Sunlight

The light of the sun can be used directly for changing chemical structures photochemically. Any industrial application must conform to the limitations imposed by the spectral distribution of the photons from the sun, the interruptions to the radiation due to the day/night rhythm, and the weather. In this review, we describe the photochemical potential of the sun, give a fundamental treatment of the concept of photoreactors driven by sunlight (abbreviated to solar photoreactors), and give an account of the realization of this concept in the first pilot plant on the “Plataforma Solar de Almeria” in southern Spain and in other activities in this field. Based on experimental data from photochemical investigations on the pilot plant scale, possibilities, limitations, and the potential growth of solar photochemistry are described. Solar photochemistry, in our opinion, is a technique which could make a contribution to the chemistry of the future because of its photochemical synthesis potential, the avoidance of waste products, and the direct utilization of the sun, not only as a primary energy source, but also as a reaction partner.     

Photochemical Cycloaromatization of Non-Benzenoid Enediynes

An efficient photo-Bergman reaction of aliphatic enediynes has been realized. Photolysis of 1 results in the formation of 4 in good yields along with [D₂] 3 . Enediyne 4 , which has never been isolated in the thermal reaction of 1 , arises here by a retro-Bergman reaction of the diradical intermediate 2 .     

Photochemical Reactions of Ligands in Transition-Metal Complexes

Three types of photochemical reaction of coordination compounds are well-known and have often been discussed: redox reactions, substitutions, and isomerizations. Less well known is a fourth type of reaction, which occurs fairly frequently: reaction of the ligands themselves. It is the purpose of this article to fill the corresponding gap in the photochemical literature and to summarize our current knowledge of such reactions and of their mechanisms and possible applications.     

Photochemical Reaction of Decacarbonyldirhenium with Thiophene and Crystal Structure of Tetradecacarbonylhydridotrirhenium

The photochemical reaction of Re₂(CO)₁₀ with thiophene in hexane solution was investigated under vacuum. Three rhenium clusters: H₂Re₃(CO)₁₂, HRe₃(CO)₁₄ and Re₃(CO)₁₄(OH)₄, were isolated. The structure of Fellmann-Kaesz cluster Complex HRe₃(CO)₁₄ was determined by use of the X-ray diffraction method. The three rhenium atoms form a plane of symmetry and L: Re₁Re₂Re₃ is 107°. The ten carbonyl groups bonded to the two terminal rhenium atoms Re₁ and Re₃, are staggered with respect to the central rhenium atoms. The bond lengths are 3.10 Å for Re₂-Re₃ and 3.34 Å for Re₁-Re₂. The bridging hydride is between Re₁ and Re₂.     

Photochemical aspects in anthracene-containing cationic polyurethanes

The synthesis, characterization and photochemical properties of a new polyurethane cationomer (PUC-AN) with anthracene chromophore groups attached on the quaternary ammonium units is reported and compared to those of a model compound (AN) carrying the same moiety. PUC-AN synthesis implied the quaternization reaction with 9-chloromethylanthracene of the tertiary amine of one precursor polymer based on poly(tetramethylene oxide) diol of 2000 average molecular weight, tolylene-2,4-diisocyanate (as 2,4- and 2,6-TDI, 80:20 v/v isomers mixture), N-methyldiethanol amine and terephthalaldehyd-bis(3-hydroxymethylphenylimine) in a molar ratio of 1:3:1:1. The fluorescence spectra of the anthracene structure indicates that the synthesized compounds present different shifts in solution and in the solid state, their photochemical behavior being influenced by monomer and excimer emission. The photophysical investigations revealed that such structures can function as fluorescent chemosensors for transitional metals, showing a fluorescence quenching in the presence of different metal ions (, Fe³⁺, Cu²⁺). The quenching mechanisms for PUC-AN and the corresponding model compound (AN) are assigned to electron transfer and/or energy transfer processes of Dexter type.     

Photochemical characterization of biomimetic molecular switches

The performance of fluorenylidene-pyrroline (FPs) and N-alkylated fluorenylidene-pyrroline (NAFPs) derivatives for their use as light-driven molecular switches has been studied. Both types of compounds share fast and controllable photoisomerization. Other competitive reaction pathways that could lead to low efficiency have been considered. Only weak fluorescence was measured and high photostability was found when irradiating these compounds for long times, together with high photoisomerization quantum yields. NAFPs are capable of using visible light, which could be useful for practical applications.     

Photochemical Processes of Superbase Generation in Xanthone Carboxylic Salts

Photobase generators are species that allow the photocatalysis of various reactions, such as thiol-Michael, thiol-isocyanate, and ring-opening polymerization reactions. However, existing compounds have complex syntheses and low quantum yields. To overcome these problems, photobase generators based on the photodecarboxylation reaction were developed. We synthesized and studied the photochemistry and photophysics of two xanthone photobase, their carboxylic acid precursors, and their photoproducts to understand the photobase generation mechanism. We determined accurate quantum yields of triplet states and photobase generation. The effect of the intermediate radical preceding the base release was demonstrated. We characterized the photophysics of the photobase by femtosecond spectroscopy and showed that the photodecarboxylation process occurred from the second excited triplet state with a rate constant of 2.2×10⁹ s⁻¹.     

Photochemical Rearrangements and Fragmentations of Alkenes and Polyenes

This article presents a systematic analysis of the numerous intramolecular reactions of electronically excited alkenes categorized according to substance class and reaction type. The rearrangements and fragmentations observed in simple alkenes are usually rediscovered in non-conjugated and conjugated dienes and polyenes, where they are accompanied by reaction types such as rearrangements involving several double bonds, cyclizations, and intramolecular cycloadditions. Geometrical factors largely determine which reaction of a series of energetically feasible competing processes actually takes place. The industrial syntheses of vitamin D utilize many of the reaction types to be discussed.     

一类酶反应底物的光化学荧光研究

研究了对位取代酚类酶反应底物的光化学荧光反应机理，指出光子不仅可以取代过氧化物酶，而且可以取代氧化剂（Ｈ_２Ｏ_２）。用于测定多种酶反应底物，结果满意。     

Enforced Electronic-Donor-Acceptor Complex Formation in Water for Photochemical Cross-Coupling

The amino alcohol meglumine solubilizes organic compounds in water and enforces the formation of electron donor acceptor (EDA) complexes of haloarenes with indoles, anilines, anisoles or thiols, which are not observed in organic solvents. UV-A photoinduced electron transfer within the EDA complexes induces the mesolytic cleavage of the halide ion and radical recombination of the arenes leading, after rearomatization and proton loss to C−C or C−S coupling products. Depending on the substitution pattern selective and unique cross-couplings are observed. UV and NMR measurements reveal the importance of the assembly for the photoinduced reaction. Enforced EDA aggregate formation in water allows new activation modes for organic photochemical synthesis.