Application In Cell Imaging Research Articles

Smartphone-Assisted and Optical Quantification of Copper and Glucose Using Palm Wine-Tailored Carbon Dots and Their Multiple Logic Gate Application.

In this work, potassium, sulfur, nitrogen, and chlorine self-doped carbon dots (CDs) were hydrothermally synthesized using palm wine as a carbon source. The palm wine-derived CDs (PW-CDs) are amorphous in nature and displayed an average particle size of 4.19 ± 0.89 nm. The as-synthesized CDs are used to fabricate a photoluminescent sensing probe to simultaneously detect Cu2+ and glucose via the "Turn ON-OFF-ON" mechanism. The PL quenching mechanism of PW-CDs enables the selective and sensitive detection of Cu2+ ions with a detection limit (LOD) of 0.8 ppb (4.7 nM). The sensing probe quantified Cu2+ in tap water, drinking water, and e-waste samples to prove its viability. Using CDs to quantify copper in e-waste leachate samples is a novel approach as no prior instances of such application have been reported. The system's performance is considered to be highly reproducible due to the relative standard deviation being <6.64%, along with excellent recoveries within the range of 93.24-109.86%. The quenched PL can be recovered by introducing glucose into the PW-CD + Cu2+ system; this strategy is employed to quantify glucose with a LOD of 0.11 ppm (0.61 μM). The feasibility of this sensor was confirmed by the determination of glucose in actual human plasma specimens of diabetic patients. It is to be noted that these samples were neither diluted nor spiked with glucose. The developed PW-CD + Cu2+ sensing system yields satisfactory recoveries of 93.45-107.37%. This probe was also incorporated into a smartphone-based sensing platform to detect Cu2+ and glucose with desirable recoveries. The proposed smartphone-based sensing platform is flexible, reliable, and accurate, making it suitable for resource-constrained areas. Furthermore, based on the effect of Cu2+ ions and glucose on the PL response and absorbance spectra of PW-CDs, four logic gates (YES, IMPLICATION, NOT, and OR) were designed, and PW-CDs were also used for cell imaging applications.

Ultrathin Boron Growth onto Nanodiamond Surfaces via Electrophilic Boron Precursors.

Diamond as a templating substrate is largely unexplored, and the unique properties of diamond, including its large bandgap, thermal conductance, and lack of cytotoxicity, makes it versatile in emergent technologies in medicine and quantum sensing. Surface termination of an inert diamond substrate and its chemical reactivity are key in generating new bonds for nucleation and growth of an overlayer material. Oxidized high-pressure high temperature (HPHT) nanodiamonds (NDs) are largely terminated by alcohols that act as nucleophiles to initiate covalent bond formation when an electrophilic reactant is available. In this work, we demonstrate a templated synthesis of ultrathin boron on ND surfaces using trigonal boron compounds. Boron trichloride (BCl3), boron tribromide (BBr3), and borane (BH3) were found to react with ND substrates at room temperature in inert conditions. BBr3 and BCl3 were highly reactive with the diamond surface, and sheet-like structures were produced and verified with electron microscopy. Surface-sensitive spectroscopies were used to probe the molecular and atomic structure of the ND constructs' surface, and quantification showed the boron shell was less than 1 nm thick after 1-24 h reactions. Observation of the reaction supports a self-terminating mechanism, similar to atomic layer deposition growth, and is likely due to the quenching of alcohols on the diamond surface. X-ray absorption spectroscopy revealed that boron-termination generated midgap electronic states that were originally predicted by density functional theory (DFT) several years ago. DFT also predicted a negative electron surface, which has yet to be confirmed experimentally here. The boron-diamond nanostructures were found to aggregate in dichloromethane and were dispersed in various solvents and characterized with dynamic light scattering for future cell imaging or cancer therapy applications using boron neutron capture therapy (BNCT). The unique templating mechanism based on nucleophilic alcohols and electrophilic trigonal precursors allows for covalent bond formation and will be of interest to researchers using diamond for quantum sensing, additive manufacturing, BNCT, and potentially as an electron emitter.

Highly Selective Drug-Derived Fluorescent Probes for the Cannabinoid Receptor Type 1 (CB1R).

The cannabinoid receptor type 1 (CB1R) is pivotal within the endocannabinoid system regulating various signaling cascades with effects in appetite regulation, pain perception, memory formation, and thermoregulation. Still, understanding of CB1R's cellular signaling, distribution, and expression dynamics is very fragmentary. Real-time visualization of CB1R is crucial for addressing these questions. Selective drug-like CB1R ligands with a defined pharmacological profile were investigated for the construction of CB1R fluorescent probes using a reverse design-approach. A modular design concept with a diethyl glycine-based building block as the centerpiece allowed for the straightforward synthesis of novel probe candidates. Validated by computational docking studies, radioligand binding, and cAMP assay, this systematic approach allowed for the identification of novel pyrrole-based CB1R fluorescent probes. Application in fluorescence-based target-engagement studies and live cell imaging exemplify the great versatility of the tailored CB1R probes for investigating CB1R localization, trafficking, pharmacology, and its pathological implications.

Unraveling the ability of electronically diverse 9H-carbazoles as potential fluorophores in cellular imaging applications with BSA binding affinities

In the present study, fluorophores based on 9H-carbazole have been synthesized from readily accessible 2-nitrochalcones in a single-step reductive cyclization reaction with methyl acetoacetate. The molecular geometry was confirmed based on IR, NMR and mass analysis. The ground state geometry for all the synthesized compounds was optimized using DFT-based methods to explore the localization of frontier molecular orbitals in electronically diverse carbazoles. The photophysical studies revealed a higher Stokes shift for PPM 17 due to the presence of the naphthyl group at C-1 of carbazole. The high stability of PPM14, as evidenced by high chemical hardness, prompted us to explore its BSA binding affinity using fluorescence titration, synchronous,3-D fluorescence and energy transfer analysis. The probe (PPM 14) displayed a “turn-off” response in the presence of Fe3+ ions. The less cytotoxic behavior of PPM 14, its “turn-off” fluorescence response with Fe3+, and its utility in cell imaging application establish it as a potential small organic framework-based bio-sensor.

A dual responsive bis-thiophene affixed thiosemicarbazide based chemosensor for colorimetrically Hg2+ and fluorometrically Cu2+ ions and their applications in live cell imaging

In this work, we developed a fast and straightforward colorimetric and photoluminescent chemosensor probe (P1), featuring bis-thiophene-thiosemicarbazide moieties as its signaling and binding unit. This probe exhibited rapid sensitivity to Hg2+ and Cu2+ ions in a semi-aqueous medium, resulting in distinct colorimetric and photoluminescent changes. In the presence of Cu2+, P1 displayed an impressive 50-fold increase in photoluminescence (PL) at 450 nm (with excitation at 365 nm). The probe P1 formed a 1:1 complex with Hg2+ and Cu2+ ions, featuring association constant values of 4.04 × 104 M−1 and 1.25 × 103 M−1, respectively. P1 has demonstrated its efficacy in the analysis of real samples, yielding promising results. Additionally, the probe successfully visualized copper ions on a mouse fibroblast cell line (NIH3T3), highlighting its potential as an intracellular probe for copper ion detection.

Highly stable, fluorescent, artificial lipoprotein nanoparticles.

Here, we report a novel kind of protein nanoparticles of 11 nm in size, which have a central protein core surrounded by two layers of lipid. One layer of the lipid was covalently attached to the protein, while the other layer has been physically assembled around the protein core. Particle synthesis is highly modular, while both the size and charge of the protein nanoparticles are controlled in a predictable manner. Circular dichroism studies of the conjugate showed that the protein secondary structure is retained, while biophysical characterizations indicated the particle purity, size, and charge. The conjugate had a high thermal stability to steam sterilization conditions at 121°C (17 psi). After labeling the protein core with few different fluorescent dyes, they were strongly fluorescent with the corresponding colors independent of their size, unlike quantum dots. They are readily digested by proteases, and these water-soluble, non-toxic, highly stable, biocompatible, and biodegradable conjugates are suitable for cell imaging and drug delivery applications.

A highly selective fluorescent sensor with aggregation-induced emission for detection of H2S and its application in live cells imaging

Hydrogen sulfide (H2S) is traditionally considered as an ecologically toxic gas, has been proven to be the third endogenous signalling gasotransmitter and plays important roles in physiological and pathophysiological conditions. Herein, we designed and synthesized a novel “turn off” fluorescent probe for the detection of hydrogen sulfide(H2S) in vitro. Probe A could detect H2S in CH3CN/PBS system with significant fluorescence quenching. Probe A also showed obviously aggregation-induced emission (AIE) effects. Fluorescence titration experiments revealed probe A combine with H2S in a 1: 1 ratio with a detection limit of 6.54 μM. Cell imaging results demonstrated the potential applicability of probe A for H2S detection in living cells. The supramolecular assembly of probe A and β-cyclodextrin further revealed the interaction with H2S.

NCS-catalyzed synthesis of 2,4,5-triaryl-1H-imidazole and its selective detection of iron ions

In this study, an efficient synthesis of 2,4,5-trisubstituted imidazoles was developed via cyclocondensation starting from o-dione, substituted aromatic aldehyde and ammonium acetate using catalyst NCS, and was characterized by HR-MS, FT-IR, 1H and 13C NMR spectroscopy. A novel fluorescence sensor 4-(1H-phenanthro[9,10-d]imidazol-2-yl)-N, N-diphenylaniline (3bd) was discovered for the detection of Fe3+ ions. The probe 3bd exhibited strong blue radiation in solution. In the presence of Fe3+ ions, a significant fluorescence change of blue light to green light was observed in the EtOH/H2O (4/1, v/v) solution of 3bd (Ag+, Al3+, Ba2+, Ca2+, Cd2+, Co2+, Cr3+, Cu2+, Fe2+, Fe3+, Hg2+, Mg2+, Mn2+, Pb2+, Sr2+, Zn2+). The limits of detection of probe 3bd for Fe3+ ions was calculated to be as low as 0.70 μM, and it had excellent anti-interference ability. The probe 3bd complexed with Fe3+ ions at 1:1, and the complexation constant was 3.93 × 104 M−1. The probe 3bd realized the detection of Fe2+ oxidation to Fe3+, and the pH showed fine stability in the environment between 4.0–7.0, which ensured its potential application prospects. It also showed good reversibility upon addition of EDTA. Furthermore, the viability of 3bd to Fe3+ had practical application in live cell imaging.

An easily accessible probe for H2S based on AIE mechanism and its application in cell imaging

Hydrogen sulfide (H2S) is a crucial endogenous gas transmitter, maintaining the redox homeostasis in biological systems. Thus, a real-time and specific H2S detection approach is urgently necessary to study the attack and management of disorders linked to H2S. In this paper, a novel fluorescent probe HMP based on AIE property for the detection of H2S was elaborately conducted. In PBS buffer solution HMP exhibited high photostability, low detection limit (0.82 μM), significant anti-interference ability and fast response time and specificity to H2S. What’s more, it could be employed for the monitoring and luminescent imaging of H2S with Hela cells, thereby broadening the prospective capacity for diagnosing associated diseases.

A novel fluorescent probe based on naphthimide for detecting Zn2+ and its application in plant imaging and cell imaging

The fluorescent probe NXZ was synthesized by introducing –OH at position 1, 8-naphthalimide 4-position and benzimidazole unit at 3-position. The Stokes shift (λem − λex) of NXZ can up to 148 nm, when Zn2+ was added to the probe solution, NXZ can quickly form the complex NXZ-Zn2+ with Zn2+ in a ratio of 1: 1 within 55 s, and then quench the fluorescence intensity by 4.4 times. The complexation constant of NXZ and Zn2+ was 1.2 × 104 M−1, and the detection limit for Zn2+ was 40.3 nM, which is much lower than the maximum Zn2+ content (76 µM) stipulated in the World Health Organization (WHO). Exploring the practical application of the probe NXZ found that NXZ can be used for the quantitative detection of Zn2+ in actual water samples and qualitatively detect Zn2+ in bean sprouts samples and Hela cells. In addition, NXZ can also be made into Zn2+ test paper.

Bifunctional fluorescent probe for fluoride and copper(Ⅱ) based on perylene diimide derivative and its application in cell imaging

A bifunctional fluorescent probe P1 based on perylene diimide derivative (PDI) was developed, demonstrating high sensitivity and selectivity for the rapid detection of fluoride (F-) and copper (Cu2+). The fluorescence of P1 was quenched upon interaction with F- and enhanced upon interaction with Cu2+. The concentration ranges for fluorescent detection of F- and Cu2+ are 0 ∼ 4 × 10-5 M and 0 ∼ 2.4 × 10-5 M, respectively. The stoichiometry of the complex formed between Cu2+ and P1 or F− and P1 was 1:1. The probe displays detection limits of 0.16 and 0.21 μM and binding constants (Ksv) of 3.92 × 104 and 1.72 × 104 M−1 for the detection of F- and Cu2+, respectively. The sensing mechanism and binding mode of P1 with F- and Cu2+ were investigated. The fluorescence quenching/enhancement of P1 was controlled by a photoinduced electron-transfer (PET) process. Furthermore, the ability of the probe to detect intracellular F- and Cu2+ was assessed using cell imaging techniques.

Parallel illumination for depletion microscopy through acousto-optic spatial light modulation

Several types of super-resolution microscopy techniques, such as Stimulated Emission Depletion (STED), Reversible Saturable Optical Fluorescence Transitions (RESOLFT) or Switching Laser Mode (SLAM) microscopies, employ Laguerre-Gaussian beams (also called vortex or doughnut beams) to obtain fluorescence information within a sub-wavelength region of the specimen under observation, thus breaking the diffraction limit and producing images of greatly improved quality. However, in general, these techniques operate on a point-by-point basis, so scanning the sample in order to build a full, meaningful image, takes time. Parallelization of the illumination is the only way to make these microscopy techniques more suitable for real-time live cell imaging applications. Here, we demonstrate the parallel production of arbitrary arrays of Gaussian and Laguerre-Gaussian lasers foci suitable for super-resolution microscopy, together with the possibility to fast scan through the sample, by means of acousto-optic spatial light modulation, a technique that we have pioneered in the past in several other fields. Parallelized illumination with both Gaussian and doughnut beams will be then used to acquire super-resolution images.

Sustainable carbon dots from Borreria hispida: enhanced colorimetric sensing of Fe3+ ions and biological applications in live cell imaging.

This study presents the synthesis of advanced nanomaterials derived from the hedge-grown herbal plant, Borreria hispida, and explores their environmental and biological applications. Using a one-step hydrothermal synthesis method, carbon dots derived from Borreria hispida (BHCD) were fabricated and thoroughly characterized through XRD, TEM, FTIR, CHNS, UV-visible, and PL spectroscopy analyses. Under UV illumination, these plant-based carbon dots demonstrated exceptional water solubility, notable photo stability, and a high quantum yield of 40.8%. The average particle size of BHCD was absorbed around 0.5 to 3.5 nm, contributing to superior selectivity and sensitivity in detecting Fe3+ ions, with a limit of detection of 1.2 × 10-6 M. Investigation into the sensing mechanism revealed a binding model wherein two carbon atom molecules bind to one Fe3+ atom in a 2 : 1 ratio for BHCDs and Fe3+ interactions. Additionally, the effectiveness of the developed fluorescent probe for Fe3+ detection was validated using real water samples from ponds and lakes, highlighting its potential for environmental monitoring applications. Furthermore, the biological effects of BHCD were evaluated through cytotoxic assays, demonstrating significant inhibitory effects on MCF7 breast cancer cell lines, with a maximum cell viability of 60%. This research underscores the multifaceted potential of BHCD in environmental monitoring and biomedical applications.

An Aggregation-Induced Fluorescence Probe for Detection H2S and Its Application in Cell Imaging.

Monitoring hydrogen sulfide (H2S) in living organisms is very important because H2S acts as a regulator in many physiological and pathological processes. Upregulation of endogenous H2S concentration has been shown to be closely related to the occurrence and development of tumors, atherosclerosis, neurodegenerative diseases and diabetes. Herin, a novel fluorescent probe HND with aggregation-induced emission was designed. Impressively, HND exhibited a high selectivity, fast response (1 min) and low detection limit (0.61 μM) for H2S in PBS buffer (10 mM, pH = 7.42). Moreover, the reaction mechanism between HND and H2S was conducted by Job's plot, HR-MS, and DFT. In particular, HND was successfully employed to detect H2S in HeLa cells.

Two-Photon Absorption Aggregation-Induced Emission Luminogen/Paclitaxel Nanoparticles for Cancer Theranostics.

Multifaceted nanoplatforms integrating fluorescence imaging and chemotherapy have garnered acknowledgment for their potential potency in cancer diagnosis and simultaneous in situ therapy. However, some drawbacks remain for traditional organic photosensitizers, such as poor photostability, short excitation wavelength, and shallow penetration depth, which will greatly lower the chemotherapy treatment efficiency. Herein, we present lipid-encapsulated two-photon active aggregation-induced emission (AIE) luminogen and paclitaxel (PTX) nanoparticles (AIE@PTX NPs) with bright red fluorescence emission, excellent photostability, and good biocompatibility. The AIE@PTX NPs exhibit dual functionality as two-photon probes for visualizing blood vessels and tumor structures, achieving penetration depth up to 186 and 120 μm, respectively. Furthermore, the tumor growth of the HeLa-xenograft model can be effectively prohibited after the fluorescence imaging-guided and PTX-induced chemotherapy, which shows great potential in the clinical application of two-photon cell and tumor fluorescence imaging and cancer treatment.

Turn-on fluorescent probe for ultrafast detection of phosgene in the environment and household bleach; its applications in live cell imaging

Phosgene is a vital industrial gas that might substantially imperil public safety in the case of an accident or terrorist attack due to its extreme toxicity and history of use as a chemical warfare agent. Herein we built a simple photoinduced electron transfer-intramolecular charge transfer based turn-on fluorescent probe for ultrafast detection of phosgene within ≤ 3 s. The probe provides low detection limit (0.01 ppm) with distinct colour and fluorescence changes only in the presence of phosgene. With the help of a readily available colour smartphone app, the probe-loaded paper strips allowed for the direct visualization and quantitative measurement of phosgene vapour. Using the probe-coated strips to detect phosgene that evolved from common household bleach. Finally, the probe was utilized to detect phosgene in living cells.

Synthesis of 1, 2, 3-triazole linked 5 fluorouracil - carbon dots -folate conjugates for target specific anticancer activity and cell imaging applications

This work provides a unique strategy to anchor the individual properties of fluorophores, pharmacophores and receptors in a single platform using a copper-catalyzed azide-alkyne cycloaddition reaction (CuAAC). Notably, this approach counters the long-term dispute associated with the target-specific drug delivery of 5 fluorouracil and its intracellular tracing. Significantly, the luminescence property of the carbon dots (CDs) and the anticancer activity of the 5 fluorouracil drug are well preserved, even after their structural modification. The resulting nano-hybrid conjugate shows good thermal stability, photo-stability and can selectively guide the drug molecule toward cancer cells and remain nontoxic to noncancerous (hFB) cells. The conjugation of folic acid to the nanohybrid surface promoted the folate receptor-facilitated endocytosis to the folate-positive (HeLa) cell lines over the folate-negative (MCF-7) cells, which enhanced cellular uptake and corresponding better cell apoptosis results. Around 18.2% of cell apoptosis (late + early) values were recorded for the folate-conjugated formulation compared to the folate-less formulation (12.3%) and pure 5 FU drug (7.9%) by a flow cytometry study. Cell cycle analysis confirmed that the populations of HeLa cells in the S phase were around 18.20% and 29.8% for the folate-less formulation and the folate-conjugated formulation, indicating all the formulations can hinder the DNA replication and thymidylate synthesis by introducing cell cycle arrest in the S phase, just like the pure 5 FU drug. Also, the location of the drug molecule can be simultaneously traced because of the luminescent nature of the CDs. Therefore, the developed system has potential in target-specific drug delivery and long-term drug molecule tracking applications.

Quantitative detection of L-cysteine with a ratiometric probe combined with smartphone imaging and fluorescent test strips

Cysteine is an important thiol involved in the vital activities of the human body, and many physiological processes in the body are associated with it. Accurate detection of cysteine is of great importance in monitoring the development of diseases. In this paper, we prepared a ratiometric flavonoid fluorescent probe, HDFA, that exhibits specific selectivity for L-cysteine. HDFA was the first fluorescent probe that can distinguish between L-cysteine and D-cysteine. When detecting L-cysteine, the fluorescence color changed from blue to yellow, and the fluorescence quantum yield increased from 16.96 % to 21.30 %. The probe has a short response time (90 s) to L-cysteine, a large Stokes shift (186 nm), good selectivity, and anti-interference, and the detection limit was 0.32 μM. The detection mechanism was confirmed by mass spectrometry. In addition, HDFA has good applications in cell imaging, smartphone fluorescence imaging and fluorescence test strip detection.

In-depth Photo-physical Properties, Characterization and Cell Imaging Applications of 2, 6-dicyanoanilines Based on Iso-phthalaldehyde and Tere-phthalaldehyde Frame

In current manuscript we have reported the interesting fluorescence properties, characterization and cell imaging applications of 2, 6-dicyanoanilines based on iso-phthalaldehyde and tere-phthalaldehyde frame. Earlier preliminary reported work was further extended to in-depth study of photo-physical properties like quantum yield, Fluorescence decay, stock shifts etc, and screening of representative compounds (1b and 2b) for use in fluorescent cell imaging applications. The 2, 6-dicyanoanilines based on iso-phthalaldehyde and tere-phthalaldehyde frame were showing very interesting photo-physical properties and promising prospectus for cell imaging applications.

A Ratiometric Fluorescent Detection Platform Using G‐CDs@[Ru(bpy)3]2+ for the Specific Detection of Hypochlorite and Live Cell Imaging

AbstractHypochlorite (ClO−), an essential reactive oxygen species (ROS) in physiological processes, is identified to be closely connected with oxidative stress and related diseases. Meanwhile, ClO− is a commonly‐used disinfector for water treatment, and in public places, under acidic conditions, it's easily decomposed into hypertoxic chlorine gas. Since the strong oxidizing property of ClO−, many oxidizing agents may disturb the ClO− detection. Specific and accurate detection of ClO− with superior sensitivity is a challenge. In this work, a sensing platform for rapid, sensitive, and specific ClO− detection is constructed using green fluorescent carbon dots (G‐CDs), with a linear detection range of 0.5–11 µm and a detection limit of 0.233 µm. Moreover, introducing a red fluorescent tripyridinium ruthenium ([Ru(bpy)3]2+) as a reference, a ratiometric fluorescence nanoprobe G‐CDs@[Ru(bpy)3]2+ is prepared and shows favorable intracellular imaging of exogenous and endogenous ClO−. With G‐CDs@[Ru(bpy)3]2+‐based test paper microarrays and a color recognition APP, a smartphone‐based sensing system for point‐of‐care testing of ClO− is also fabricated. In summary, this work proposed a versatile and economical smartphone‐based sensing system that featured reliability and simplicity, and suggested its potential applications in environmental water quality monitoring and live cell imaging.