Chemical Control of Fluorescence Lifetime towards Multiplexing Imaging.

Fluorescence lifetime imaging has been a powerful tool for biomedical research. Recently, fluorescence lifetime-based multiplexing imaging has expanded imaging channels by using probes that harbor the same spectral channels and distinct excited state lifetime. While it is desirable to control the excited state lifetime of any given fluorescent probes, the rational control of fluorescence lifetimes remains a challenge. Herein, we chose boron dipyrromethene (BODIPY) as a model system and provided chemical strategies to regulate the fluorescence lifetime of its derivatives with varying spectral features. We find electronegativity of structural substituents at the 8' and 5' positions is important to control the lifetime for the green-emitting and red-emitting BODIPY scaffolds. Mechanistically, such influences are exerted via the photo-induced electron transfer and the intramolecular charge transfer processes for the 8' and 5' positions of BODIPY, respectively. Based on these principles, we have generated a group of BODIPY probes that enable imaging experiments to separate multiple targets using fluorescence lifetime as a signal. In addition to BODIPY, we envision modulation of electronegativity of chemical substituents could serve as a feasible strategy to achieve rational control of fluorescence lifetime for a variety of small molecule fluorophores.

Highly Efficient Red/NIR-Emissive Fluorescent Probe with Polarity-Sensitive Character for Visualizing Cellular Lipid Droplets and Determining Their Polarity.

Lipid droplets (LDs), which are ubiquitous organelles existing in almost all eukaryotic cells, have attracted a lot of attention in the field of cell biology over the last decade. For the biological study of LDs via fluorescence imaging, the superior LD fluorescent probes with environmental polarity-sensitive character are highly desired and powerful but are very scarce. Herein, we have newly developed such a kind of fluorescent probe named LDs-Red which enables us to visualize LDs and to further reveal their polarity information. This fluorescent probe displays the advantages of intense red/near-infrared emission, high LD staining specificity, and good photostability; thus, it would be very useful for LD fluorescence imaging application. As a result, the three-dimensional confocal imaging to visualize spatial distribution of LDs and the multicolor confocal imaging to simultaneously observe LDs and other cellular organelles have been realized using this new LD fluorescent probe. Furthermore, the polarity-sensitive emission character of this probe enables us to quantitatively determine the LD polarity via spectral scan imaging. Consequently, the cancer cells (HepG2, HeLa, and Panc02) displaying lower polarity of LDs than the normal cells (L929, U251, and HT22) have been systematically demonstrated. In addition, this polarity-sensitive probe displaying shorter fluorescence wavelengths in cancer cells than in normal cells has an important and potential ability to distinguish them.

STED Nanoscopy Imaging of Cellular Lipid Droplets Employing a Superior Organic Fluorescent Probe.

Lipid droplets (LDs) are spherical organelles that participate in numerous biological processes. In order to visualize LDs on the nanoscale, nanoscopy fluorescence imaging is considered as the most attractive technique but is substantially limited by the characteristics of fluorescent probes. Thus, the development of a superior fluorescent probe that is capable of nanoscopy fluorescence imaging has attracted enormous attention. Herein, a benzodithiophene-tetraoxide-based molecule Lipi-BDTO has been developed that can easily undergo the stimulated emission depletion (STED) process and displays high photostability. These two characteristics of fluorescent probes finely satisfy the requirements of STED nanoscopy imaging. Indeed, applying the probe for STED imaging achieves a high resolution of 65 nm, belonging to one of the leading results of LDs fluorescence imaging. Furthermore, the high photostability of this fluorescent probe enables it to monitor the dynamics of LDs by time-lapse STED imaging as well as to visualize the three-dimensional (3D) spatial distribution of LDs by 3D STED imaging. Notably, the resolution of the 3D STED image represents one of the best LDs fluorescence imaging results so far. Besides STED nanoscopy imaging, the superior utility of this fluorescent probe has been also demonstrated in two-color 3D confocal imaging and four-color confocal imaging.

Modal perspective on geometric parametric instability sidebands in graded-index multimode fibers.

In this paper, we investigated the geometric parametric instability (GPI) in graded-index multimode fibers through the multimode generalized nonlinear Schrödinger equation. Our results clearly and intuitively indicate that the generations of GPI sidebands are nearly synchronous in the spectrums of all modes, and the shapes of these spectrums are nearly the same. The numerical results show that the energies of the GPI sidebands come from the pump sideband, and these sidebands are carried by similar spatial beam profiles due to the similar modal components. We also found that the large modal dispersion has an influence for the symmetry of these GPI sidebands.

Self-Healing, Laminated, and Low Resistance NH3 Sensor Based on 6,6',6″-(Nitrilotris(benzene-4,1-diyl))tris(5-phenylpyrazine-2,3-dicarbonitrile) Sensing Material Operating at Room Temperature.

With the rapid development of the concept of the Internet of Things (IoT), gas sensors with the function of simulating the human sense of smell became irreplaceable as a key element. Among them, ammonia (NH3) sensors played an important role in respiration tests, environmental monitoring, safety, and other fields. However, the fabrication of the high-performance device with high stability and resistance to mechanical damages was still a challenge. In this work, polyurethane (PU) with excellent self-healing ability was applied as the substrate, and the sensor was designed from new sensitive material design and device structure optimization, through applying the organic molecule with groups which could absorb NH3 and the laminated structure to shorten the electronic transmission path to achieve a low resistance state and favorable sensing properties. Accordingly, a room temperature flexible NH3 sensor based on 6,6',6″-(nitrilotris(benzene-4,1-diyl))tris(5-phenylpyrazine-2,3-dicarbonitrile) (TPA-3DCNPZ) was successfully developed. The device could self-heal by means of a thermal evaporation assisted method. It exhibited a detection limit of 1 ppm at 98% relative humidity (RH), as well as great stability, selectivity, bending flexibility, and self-healing properties. The improved NH3 sensing performance under high RH was further investigated by complex impedance plots (CIPs) and density functional theory (DFT), attributing to the enhanced adsorption of NH3. The TPA-3DCNPZ based NH3 sensors proved to have great potential for application on simulated exhaled breath to determine the severity of kidney diseases and the progress of treatment. This work also provided new ideas for the construction of high-performance room temperature NH3 sensors.

Fast degradation of macro alkanes through activating indigenous bacteria using biosurfactants produced by Burkholderia sp.

Soil bacteria that produce biosurfactants can use total petroleum hydrocarbons (TPHs) as a carbon source. This study demonstrated that biosurfactants produced by Burkholderia sp. enhanced the recovery and synergism of soil microbial community, resulting in fast degradation of macro alkanes. Experiments were carried out by applying bio-stimulation after pre-oxidation to investigate the effects of nutrient addition on biosurfactant production, TPH degradation, and microbial community succession in the soil. The results presented that bio-stimulation could produce biosurfactants in high C/N (32.6) and C/H (13.3) conversion after pre-oxidation and increased the total removal rate of TPH (10.59-46.71%). The number of total bacteria had a rapid increase trend (2.94-8.50 Log CFU/g soil). The degradation rates of macro alkanes showed a 4.0-fold (48.07 mg/kg·d-1 versus 186.48 mg/kg·d-1) increase, and the bioremediation time of degrading macro alkanes saved 166 days. Further characterization revealed that the biosurfactants produced by Burkholderia sp. could activate indigenous bacteria to degrade macro alkanes rapidly. A shift in phylum from Actinomycetes to Proteobacteria was observed during bioremediation. The average relative abundance of the microbial community increased from 36.24 to 64.96%, and the predominant genus tended to convert from Allorhizobium (8.57%) to Burkholderia (15.95%) and Bacillus (15.70%). The co-occurrence network and Pearson correlation analysis suggested that the synergism of microbial community was the main reason for the fast degradation of macro alkanes in petroleum-contaminated soils. Overall, this study indicated the potential of the biosurfactants to activate and enhance the recovery of indigenous bacteria after pre-oxidation, which was an effective method to remediate petroleum-contaminated soils.

Construction of a red emission fluorescent probe for selectively detection of cysteine in living cells.

Cysteine (Cys) is a vital amino acid in the body, and its abnormal expression level is associated with many diseases. In this study, a novel fluorescent probe ACHB was synthesized, showing high selectivity, anti-interference ability and achieving accurate detection of cysteine. Different from most previous off-on probes, ACHB showed an on-off fluorescence response to Cys. Acrylic ester was used as a recognizer while green fluorescence protein (GFP) chromophore derivative 4-hydroxybenzylidene-imidazolinone (HBI) was used as the fluorophore. The addition of Cys leads to the hydrolysis of the red-emitting probe (613 nm), releasing a precursor with a lower fluorescent signal and showing an on-off spectral signal, which was ideal for obtaining sensitive detection with high specificity. Furthermore, the probe was successfully applied for simultaneous determination of cysteine (Cys) in living cells and biological sample (mouse serum). In conclusion, probe ACHB is a promising tool to display the intracellular cysteine concentration level, providing a good visualization method for clinical diagnosis and scientific basic research.

Super-resolution dynamic tracking of cellular lipid droplets employing with a photostable deep red fluorogenic probe.

Lipid droplets (LDs) are critical organelles involved in many physiological processes in eukaryotic cells. To visualize and study LDs, particular the small/nascent LDs, the emerging super-resolution fluorescence imaging techniques with nanoscale resolution would be much more powerful in comparison to the conventional confocal/wide-field imaging techniques. However, directly limited by the availability of advanced LDs probes, super-resolution fluorescence imaging of LDs is a practically challenging task. In this context, a superior LDs fluorescent probe named Lipi-Deep Red is newly developed for structured illumination microscopy (SIM) super-resolution imaging. This fluorescent probe features with the advantages of strong deep red/NIR emission, fluorogenic character, high LDs specificity, and outstanding photostability. These advantages enable the fluorescent probe to be finely applied in SIM super-resolution imaging, e.g. time-lapse imaging (up to 1000 frames) to monitor the LDs dynamics at nanoscale (159 nm), two-color time-lapse imaging to discover the nearby contact/interaction between LDs and mitochondria. Consequently, the fusion processes of LDs are impressively visualized at a high spatial and temporal resolution. Two kinds of contact models between LDs and mitochondria (dynamic contact and stable contact) newly proposed in the recent literatures are successfully revealed.

Ultrabright organic fluorescent probe for quantifying the dynamics of cytosolic/nuclear lipid droplets.

Lipid droplets (LDs) are extremely active organelles that play a crucial role in energy metabolism, membrane formation, and the production of lipid-derived signaling molecules by regulating lipid storage and release. Nevertheless, directly limited by the lack of superior fluorescent probes, studies of LDs dynamic motion velocity have been rarely reported, especially for nuclear LDs. Herein, a novel organic fluorescent probe Lipi-Bright has been rationally developed based on bridged cyclization of distyrylbenzene. The fully ring-fused molecule structure endows the probe with high photostability. Moreover, this new fluorescent probe displays the features of excellent LDs staining specificity as well as ultrahigh fluorescence brightness. Lipi-Bright labeled LDs was dozens of times brighter than representative probes BODIPY 493/503 or Nile Red. Consequently, by in-situ time-lapse fluorescence imaging, the dynamics of LDs have been quantitatively studied. For instance, the velocities of cytosolic LDs (37 ± 15 nm/s) are found to be obviously faster than those of nuclear LDs (24 ± 4 nm/s), and both the cytosolic LDs and the nuclear LDs would be moved faster or slower depend on the various stimulations. Overall, this work providing plentiful information on LDs dynamics will greatly facilitate the in-depth investigation of lipid metabolism.

Molecular Conformation Engineering To Achieve Longer and Brighter Deep Red/Near-Infrared Emission in Crystalline State.

A series of molecules 1-5 containing the same fluorophore and different alkyl chains are synthesized to reveal the significant effect of molecular conformations on the emission properties. In crystalline state, molecules 1-3 exhibit strong orange emissions with maxima (λem) of about 600 nm and quantum yields (ΦF) of around 60%, while molecules 4 and 5 display much longer emissions to the deep red/near-infrared (NIR) region as well as even higher efficiencies (λem = 693 nm, ΦF = 73% for 4; λem = 654 nm, ΦF = 93% for 5). The largely red-shifted emissions of 4 and 5 as well as the significantly improved ΦF are very unusual. Furthermore, the ΦF of 4 and 5 represent the highest values among organic solids with similar deep red/NIR emission wavelengths. On the basis of the experimental measurements and theoretical calculations, the new molecular design of conformation engineering, the impressive emission properties, and the potential NIR fluorescence sensing and lasing applications are comprehensively investigated.

Lysosome targeting metal-organic framework probe LysFP@ZIF-8 for highly sensitive quantification of carboxylesterase 1 and organophosphates in living cells.

Herein, a lysosomal targeting LysFP@ZIF-8 metal-organic framework (MOF) was fabricated using fluorescent protein chromophore-based probe (LysFP) for selectively detection of carboxylesterase 1 (CES1) in living cells. Unlike the regular small molecule fluorescent probes, LysFP@ZIF-8 showed wide range pH tolerabiligy, high selectivity and sensitivity to CES1 in bio-samples, and was successfully applied to achieve the visual monitoring of CES1 activity in living cells. Low detection limit and high fluorescence quantum yield was calculated as 79 ng/mL and 0.76 for LysFP@ZIF-8, respectively. Furthermore, LysFP@ZIF-8 can also serve as a fluorescence indicator of organophosphates pesticide exposure in the way of hydrolyzing the carboxylic acid ester group in LysFP. This type of probe can inspire the development of fluorescent tools for further explore many pathological processes.

Detection of Carboxylesterase 1 and Chlorpyrifos with ZIF-8 Metal-Organic Frameworks Using a Red Emission BODIPY-Based Probe.

In this work, a red emission fluorescent probe CBZ-BOD@zeolitic imidazolate framework-8 (ZIF-8) was fabricated based on metal-organic frameworks (MOFs) for detecting carboxylesterase 1 (CES1). The small molecule probe CBZ-BOD was first synthesized and then used to prepare the functionalized MOF material. ZIF-8 was chosen as an encapsulation shell to improve the detection properties of CBZ-BOD. Using this unique porous materials, ultrasensitive quantification of CES1 and chlorpyrifos was successfully realized. The low detection limit and high fluorescence quantum yield were calculated as 1.15 ng/mL and 0.65 for CBZ-BOD@ZIF-8, respectively. CBZ-BOD@ZIF-8 has good biocompatibility and was successfully applied to monitor the activity of CES1 in living cells. A molecular docking study was used to explore the binding of CES1 and CBZ-BOD, finding that CES1 can bind with the probe before and after hydrolysis. This type of materialized probe can inspire the development of fluorescent tools for further exploration of many pathological processes.

Construction of a red emission fluorescent protein chromophore-based probe for detection of carboxylesterase 1 and carbamate pesticide in culture cells.

Designing fluorescent probe for detecting carboxylesterase 1 is remains challenging. Herein, a red emission human carboxylesterase 1 (CES1) probe (CAE-FP) was synthesized based on fluorescent protein chromophore. Probe CAE-FP can specific detect human CES1 with high selectively. The fluorescence quantum yield was calucated as 0.19. The carboxylic acid ester in CAE-FP could be easily hydrolyzed by CES1 under physiological conditions, and this process could induce the obvious fluorescence signal in red emission region. The detection limit of CES1 was calculated as 84.5 ng/mL. Due to the biological detoxification mechanism of carboxylesterase and the obvious inhibitory effect of pesticides on its activity, CAE-FP was applied to detect carbamate pesticide and have achieved good application results. Since fluorescent protein chromophore has excellent biocompatibility, probe CAE-FP with good cell membrane permeable and was successfully applied to monitor the real activities of CES1 in living cells. In summary, this is one of the few reported fluorescent probes that can specific detect the real-time activity of CES1 in biological samples. Besides, we first applied the fluorescent protein chromophore to construct the specific target enzyme probe. This work would contribute to further investigate CES1-associated physiological and pathological processe.

Numerical and Theoretical Study of Tunable Plasmonically Induced Transparency Effect Based on Bright-Dark Mode Coupling in Graphene Metasurface.

In this paper, we numerically and theoretically study the tunable plasmonically induced transparency (PIT) effect based on the graphene metasurface structure consisting of a graphene cut wire (CW) resonator and double split-ring resonators (SRRs) in the middle infrared region (MIR). Both the theoretical calculations according to the coupled harmonic oscillator model and simulation results indicate that the realization of the PIT effect significantly depends on the coupling distance and the coupling strength between the CW resonator and SRRs. In addition, the geometrical parameters of the CW resonator and the number of the graphene layers can alter the optical response of the graphene structure. Particularly, compared with the metal-based metamaterial, the PIT effect realized in the proposed metasurface can be flexibly modulated without adding other actively controlled materials and reconstructing the structure by taking advantage of the tunable complex surface conductivity of the graphene. These results could find significant applications in ultrafast variable optical attenuators, sensors and slow light devices.

A minireview of viscosity-sensitive fluorescent probes: design and biological applications.

Microenvironment-related parameters like viscosity, polarity, and pH play important roles in controlling the physical or chemical behaviors of local molecules, which determine the physical or chemical behaviors of surrounding molecules. In general, changes of the internal microenvironment will usually lead to cellular malfunction or the occurrence of relevant diseases. In the last few decades, the field of chemicobiology has received great attention. Also, remarkable progress has been made in developing viscosity-sensitive fluorescent probes. These probes were particularly efficient for imaging viscosity in biomembranes as well as lighting up specific organelles, such as mitochondria and lysosome. Besides, there are some fluorescent probes that can be used to quantify intracellular viscosity when combined with fluorescence lifetime (FLIM) and ratiometric imaging under water-free conditions. In this review, we summarized the majority of viscosity-sensitive chemosensors that have been reported thus far.