Membrane potential is a fundamental property of biological cells. Changes in membrane potential characterize a vast number of vital biological processes, such as the activity of neurons and ...cardiomyocytes, tumorogenesis, cell-cycle progression, etc. A common strategy to record membrane potential changes that occur in the process of interest is to utilize organic dyes or genetically-encoded voltage indicators with voltage-dependent fluorescence. Sensors are introduced into target cells, and alterations of fluorescence intensity are recorded with optical methods. Techniques that allow recording relative changes of membrane potential and do not take into account fluorescence alterations due to factors other than membrane voltage are already widely used in modern biological and biomedical studies. Such techniques have been reviewed previously in many works. However, in order to investigate a number of processes, especially long-term processes, the measured signal must be corrected to exclude the contribution from voltage-independent factors or even absolute values of cell membrane potential have to be evaluated. Techniques that enable such measurements are the subject of this review.
Ultrafast transient absorption spectroscopy was used to investigate the nature of photoinduced charge transfer processes taking place in ZnSe/CdS/Pt colloidal heteronanocrystals. These nanoparticles ...consist of a dot-in-a-rod semiconductor domain (ZnSe/CdS) coupled to a Pt tip. Together the three components are designed to dissociate an electron–hole pair by pinning the hole in the ZnSe domain while allowing the electron to transfer into the Pt tip. Separated charges can then induce a catalytic reaction, such as the light-driven hydrogen production. Present measurements demonstrate that the internal electron–hole separation is fast and results in the localization of both charges in nonadjacent parts of the nanoparticle. In particular, we show that photoinduced holes become confined within the ZnSe domain in less than 2 ps, while electrons take approximately 15 ps to transition into a Pt tip. More importantly, we demonstrate that the presence of the ZnSe dot within the CdS nanorods plays a key role both in enabling photoinduced separation of charges and in suppressing their backward recombination. The implications of the observed exciton dynamics to photocatalytic function of ZnSe/CdS/Pt heteronanocrystals are discussed.
A simple approach for in situ laser-induced modification of iridium-based materials to increase their electrocatalytic activity towards enzyme-free glucose sensing was proposed. For this purpose, we ...deposited gold and platinum separately and as a mixture on the surface of pre-synthesized iridium microstructures upon laser irradiation at a wavelength of 532 nm. Then, we carried out the comparative investigation of their morphology, elemental and phase composition as well as their electrochemical properties. The best morphology and, as a result, the highest sensitivity (~9960 µA/mM cm2) with respect to non-enzymatic determination of D-glucose were demonstrated by iridium-gold-platinum microstructures also showing low limit of detection (~0.12 µM), a wide linear range (0.5 µM–1 mM) along with good selectivity, reproducibility and stability.
In this work, the method of selective laser reductive sintering was used to fabricate the sensor-active copper and nickel microstructures on the surface of glass-ceramics suitable for non-enzymatic ...detection of glucose. The calculated sensitivities for these microsensors are 1110 and 2080 μA mM−1·cm−2 for copper and nickel, respectively. Linear regime of enzymeless glucose sensing is provided between 0.003 and 3 mM for copper and between 0.01 and 3 mM for nickel. Limits of glucose detection for these manufactured micropatterns are equal to 0.91 and 2.1 µM for copper and nickel, respectively. In addition, the fabricated materials demonstrate rather good selectivity, long-term stability and reproducibility.
Fluorescence of the vast majority of natural opsin-based photoactive proteins is extremely low, in accordance with their functions that depend on efficient transduction of absorbed light energy. ...However, several recently proposed classes of engineered rhodopsins with enhanced fluorescence, along with the discovery of a new natural highly fluorescent rhodopsin, NeoR, opened a way to exploit these transmembrane proteins as fluorescent sensors and draw more attention to studies on this untypical rhodopsin property. Here, we review the available data on the fluorescence of the retinal chromophore in microbial and animal rhodopsins and their photocycle intermediates, as well as different isomers of the protonated retinal Schiff base in various solvents and the gas phase.
Quantum mechanics/molecular mechanics (QM/MM) models are a widely used tool to obtain detailed insight into the properties and functioning of proteins. The outcome of QM/MM studies heavily depends on ...the quality of the applied QM/MM model. Prediction and right placement of internal water molecules in protein cavities is one of the critical parts of any QM/MM model construction. Herein, we performed a systematic study of four protein hydration algorithms. We tested these algorithms for their ability to predict X-ray-resolved water molecules for a set of membrane photosensitive rhodopsin proteins, as well as the influence of the applied water placement algorithms on the QM/MM calculated absorption maxima (
λ
max
) of these proteins. We used 49 rhodopsins and their intermediates with available X-ray structures as the test set. We found that a proper choice of hydration algorithms and setups is needed to predict functionally important water molecules in the chromophore-binding cavity of rhodopsins, such as the water cluster in the N-H region of bacteriorhodopsin or two water molecules in the binding pocket of bovine visual rhodopsin. The QM/MM calculated
λ
max
of rhodopsins is also quite sensitive to the applied protein hydration protocols. The best methodology allows obtaining an 18.0 nm average value for the absolute deviation of the calculated
λ
max
from the experimental
λ
max
. Although the major effect of water molecules on
λ
max
originates from the water molecules located in the binding pocket, the water molecules outside the binding pocket also affect the calculated
λ
max
mainly by causing a reorganization of the protein structure. The results reported in this study can be used for the evaluation and further development of hydration methodologies, in general, and rhodopsin QM/MM models, in particular.
Accurate prediction of water molecules in protein cavities is an important factor for obtaining high-quality rhodopsin QM/MM models.
In the current study, the method of Selective Surface Activation Induced by Laser (SSAIL) was used for the fabrication of metallic and bimetallic structures based on copper and gold on the surface of ...glass and glass-ceramics. It was shown that the fabricated electrodes are suitable for non-enzymatic detection of biologically essential analytes such as glucose. The implemented approach allows performing high-rate metallization of various dielectrics. Voltammetric methods were applied to evaluate the electrocatalytic activity of the obtained structures, which were used as working electrodes. The most promising results were revealed by copper-gold electrode structures manufactured on glass-ceramics. For these structures, sensitivity towards glucose sensing was 3060 μA mM
−1
cm
−2
. The linear range of glucose detection varied between 0.3 and 1000 μM. Besides, the manufactured electrodes exhibited high selectivity and long-term stability.
In the current study, the method of Selective Surface Activation Induced by Laser (SSAIL) was used for the fabrication of metallic and bimetallic structures based on copper and gold on the surface of glass and glass-ceramics.
We report one-step in situ laser-induced synthesis of the conductive copper microstructures doped with iron, zinc, nickel, and cobalt with highly developed surface area. It was observed that the ...presence of chlorides of the aforementioned metals in the solutions used in our experiments increases the deposition rate and the amount of copper in the resulting deposits; it also leads to the deposit miniaturization. The laser deposition from solutions containing cobalt (II) chloride of concentration more than 0.003 M results in fabrication of copper microelectrode with better electrochemical properties than those deposited from solutions containing chlorides of other metals of the same concentration. Moreover, copper microelectrode doped with cobalt has demonstrated good reproducibility and long-run stability as well as sensitivity and selectivity towards determination of hydrogen peroxide (limit of detection-0.2 μM) and d-glucose (limit of detection-2.2 μM). Thus, in this article we have shown the opportunity to manufacture two-phase microcomposite materials with good electrical conductivity and electrochemical characteristics using in situ laser-induced metal deposition technique. These materials might be quite useful in development of new perspective sensors for non-enzymatic detection of such important analytes as hydrogen peroxide and glucose.
Display omitted
•We propose one-step laser-induced synthesis of bimetallic microcomposite sensors.•Doping with iron, zinc, nickel, and cobalt decreases the copper electrode size.•Doping with iron, zinc, nickel, and cobalt increases the copper deposition rate.•Copper doped with cobalt reveals the best electrochemical and sensor properties.
A typical feature of proteins from the rhodopsin family is the sensitivity of their absorption band maximum to protein amino acid composition. For this reason, studies of these proteins often require ...methodologies that determine spectral shift caused by amino acid substitutions. Generally, quantum mechanics/molecular mechanics models allow for the calculation of a substitution-induced spectral shift with high accuracy, but their application is not always easy and requires special knowledge. In the present study, we propose simple models that allow us to estimate the direct effect of a charged or polar residue substitution without extensive calculations using only rhodopsin three-dimensional structure and plots or tables that are provided in this article. The models are based on absorption maximum values calculated at the SORCI+Q level of theory for cis- and trans-forms of retinal protonated Schiff base in an external electrostatic field of charges and dipoles. Each value corresponds to a certain position of a charged or polar residue relative to the retinal chromophore. The proposed approach was evaluated against an example set consisting of twelve bovine rhodopsin and sodium pumping rhodopsin mutants. The limits of the applicability of the models are also discussed. The results of our study can be useful for the interpretation of experimental data and for the rational design of rhodopsins with required spectral properties.
Cu-catalyzed 1,3-dipolar cycloaddition of ethyl 2-azidoacetate to iodobuta-1,3-diynes and subsequent Sonogashira cross-coupling were used to synthesize a large series of new triazole-based push-pull ...chromophores: 4,5-bis(arylethynyl)-1
-1,2,3-triazoles. The study of their optical properties revealed that all molecules have fluorescence properties, the Stokes shift values of which exceed 150 nm. The fluorescent properties of triazoles are easily adjustable depending on the nature of the substituents attached to aryl rings of the arylethynyl moieties at the C4 and C5 atoms of the triazole core. The possibility of 4,5-bis(arylethynyl)-1,2,3-triazoles' application for labeling was demonstrated using proteins and the HEK293 cell line. The results of an MTT test on two distinct cell lines, HEK293 and HeLa, revealed the low cytotoxicity of 4,5-bis(arylethynyl)triazoles, which makes them promising fluorescent tags for labeling and tracking biomolecules.