pH-sensitive Molecules Research Articles

Green composite synthesis based on aluminum fumarate metal-organic framework (MOF) for doxorubicin delivery: An in vitro study

The development of an eco-friendly and cost-effective drug delivery system (DDS) remains a challenge in cancer treatment. This study suggests a novel, eco-friendly, and cost-effective method to synthesize aluminum fumarate (A520) MOF using tangerine peel extract (TPE) as a pH-sensitive coating agent on a reduced graphene oxide (rGO) substrate. The system also incorporates the 2,2′-bipyridine dimethyltin diisothiocyanate complex, (Sn(bpy), as an organotin complex) to enhance therapeutic efficacy while minimizing off-target effects. In vitro cellular and molecular cytotoxicity examinations were conducted using HEK-293 and MCF-7 cell lines to evaluate cytotoxicity after 24 and 72 h. The findings of the MTT assay indicated that the final nanocarriers, without and with doxorubicin (DOX), exhibited relative cell viability of 95 % and 55 %, respectively, highlighting the system's potential for efficient and safe drug delivery. The TPE coating created a pH-sensitive DDS with high drug loading efficiency (DLE) (59.1 %) and low cytotoxicity. Drug release was studied over 200 h at pH 5.5 and 7.2. The augmented drug release profile (47 %) demonstrated a targeted release and successful inhibition of premature release in acidic environments due to pH-sensitive TPE molecules. The efficacy of targeted drug delivery to the MCF-7 cell was evaluated using 2D fluorescence microscopy, which demonstrated successful delivery of drugs to the cells. This innovative approach addresses the limitations of existing studies while promoting biocompatibility, stability, and functionality in DDSs.

Probing the Catalyst-Ionomer Interface during Electrochemical CO2 Reduction

The catalyst-ionomer interface can significantly impact the selectivity of the electrochemical CO2 reduction (CO2R) reaction. Ionomers such as Nafion are commonly used in conjunction with nanostructured CO2R catalysts in flow cells and zero-gap devices, with their presumed function being to act as a binder for the catalyst. However, recent studies have reported that ionomers such as Nafion and Sustainion can affect the reaction rate and selectivity of CO2R, and the roles of ionomer in controlling mass transfer and local pH have been hypothesized. We employed in situ Raman spectroscopy to examine CO2R intermediate and local pH on a sputtered copper (sCu) surface with and without an ionomer overlayer. We observed a difference in the potential-dependent formation of surface CO in the presence of Nafion compared to clean sCu. Using a combination of pH-sensitive probe molecules, we also determined the pH at the catalyst-ionomer interface.

Design, synthesis and application of polarity- and pH-sensitive dual-responsive small molecule fluorescent probe for early cancer diagnosis

Cancer is one of the deadliest and most complex diseases globally. It is well known that cancer cells exhibit a unique intracellular microenvironment characterized by low pH and low polarity compared to normal cells. Thus, dual-responsive fluorescent probe are more suitable for such complex and variable physiological environments than single-response probe. In this study, we designed and synthesized a dual-responsive fluorescent probe, CCT, which responds to both pH and polarity based on intramolecular charge transfer (ICT) and photoelectron transfer (PET) mechanisms.We synthesized probe CCT by combining naphthylimide, a coumarin unit, and morpholine through a six-step process. The PET mechanism between morpholine and naphthylimide renders probe CCT pH-sensitive. Additionally, the direct linkage of the polarity-sensitive units, coumarin and naphthylimide, creates a larger polarity-sensitive system. When both low pH and low polarity conditions are present, the fluorescence intensity of probe CCT is significantly higher than under separate conditions. This increased fluorescence intensity matches the low pH and low polarity microenvironment of cancer cells, facilitating bioimaging.Fluorescence imaging results demonstrate that probe CCT effectively distinguishes between cancer and normal cells, suggesting its potential as a powerful tool for early cancer diagnosis.

Rapid intracellular pH measurement based on electroporation- surface-enhanced Raman scattering

In this study, electroporation-surface-enhanced Raman scattering (SERS) was applied to rapidly measure intracellular pH. The generation of a sensitive SERS probe for measuring pH in the range of 6.0–8.0 was accomplished through the conjugation of the pH-sensitive molecule 4-mercaptobenzoic acid (4-MBA) to the surface of gold nanoparticles (Au NPs) through its thiol functional group. This bioprobe was then rapidly introduced into nasopharyngeal carcinoma CNE-1 cells by electroporation, followed by SERS scanning and the fitting of intensity ratios of each detection point’s Raman peaks at 1423 cm−1 and 1072 cm−1, to create the pH distribution map of CNE-1 cells. The electroporation-SERS assay introduces pH bioprobes into a living cell in a very short time and disperses the nanoprobe throughout the cytoplasm, ultimately enabling rapid and comprehensive pH analysis of the entire cell. Our work demonstrates the potential of electroporation-SERS for the biochemical analysis of live cells.

Chaotic-flow-driven mixing in T- and V-shaped micromixers

T- and V-shaped (or arrow-shaped) micromixers enable the control of rapid chemical reactions by permitting accelerated mixing. Their mixing performance has been extensively investigated at moderate flow rates, which correspond to Reynolds numbers of the order of several hundreds. However, this operating range does not align with the recent applications of microreactors in organic synthesis. Therefore, this study examined mixing times in T- and V-shaped micromixers with 250-µm-wide channels at high flow rates (Reynolds numbers of up to 1500). The progress of mixing between an acidic solution and a basic solution containing a pH-sensitive fluorescent molecule was visualized by fluorescence microscopy imaging. An image analysis of 200 frames for each condition helped evaluate the degree of mixing and its stability over time under chaotic flow. The performance and stability of the V-shaped micromixer were significantly enhanced when the flow-rate ratio was changed from 1:1 to 1:2. The difference in momentum between the two inlet streams suppressed the undesirable alternation of the flow patterns at the junction. The effects of the choice of fluids (in the lower- and higher-flow-rate sides) on the reaction selectivity were explored through a numerical analysis. It was clarified that the substrate-side flow rate has to be set lower than that of the reactant side in the competing consecutive reaction system. Moreover, a short mixing time of 1 ms was achieved using the V-shaped micromixer at a flow-rate ratio of 1:2 and Re of 1500. These findings provide practical guidelines for the design and operation of microreactor systems.

Halochromic Silk Fabric as a Reversible pH-Sensor Based on a Novel 2-Aminoimidazole Azo Dye

Textiles are important components for the development of lightweight and flexible displays useful in smart materials. In particular, halochromic textiles are fibrous materials with a color-changing ability triggered by pH variations mainly based on pH-sensitive dye molecules. Recently, a novel class of 2-aminoimidazole azo dyes was developed with distinct substituent patterns. In this work, silk fabric was functionalized through exhaustion for the first time with one of these dyes (AzoIz.Pip). The halochromic properties of the dye were assessed in an aqueous solution and after silk functionalization. The solutions and the fabrics were thoroughly analyzed by ultraviolet-visible (UV-vis) spectra, color strength (K/S), color difference (∆E), CIE L*a*b* coordinates, and the ultraviolet protection factor (UPF). The dyeing process was optimized, and the halochromic performance (and reversibility) was assessed in universal Britton-Robinson buffers (ranging from pH 3 to 12) and artificial body fluids (acid and alkaline perspiration, and wound exudate). AzoIz.Pip showed vibrant colors and attractive halochromic properties with a hypsochromic shift from blue (557 nm) to magenta (536 nm) in aqueous buffered solutions. Similarly, the functionalized silk showed a shift in wavelength of the maximum K/S value from 590 nm to 560 nm when pH increases. The silk fabric showed a high affinity to AzoIz.Pip, and promoted additional color stabilization of the dye, avoiding color loss as observed when the dye is in solution at alkaline pH after 24 h. The color reversibility was effective up to the fourth cycle and the fastness tests denoted suitable results, except washing fastness. The cytotoxicity of the silk fabric extracts was assessed, depicting reduced viability of HaCaT cells to <70% only when the dye concentration in the fabric is higher or equal to 64 μg·mL-1. Nevertheless, lower concentrations were also very effective for the halochromic performance in silk. These materials can thus be a helpful tool for developing sensors in several sectors such as biomedicine, packaging, filtration, agriculture, protective apparel, sports, camouflage, architecture, and design.

Coarse-Grained Molecular Dynamics of pH-Sensitive Lipids.

pH-sensitive lipids represent a class of lipids that can be protonated and destabilized in acidic environments, as they become positively charged in response to low-pH conditions. They can be incorporated into lipidic nanoparticles such as liposomes, which are able to change their properties and allow specific drug delivery at the acidic conditions encountered in some pathological microenvironments. In this work, we used coarse-grained molecular-dynamic simulations to study the stability of neutral and charged lipid bilayers containing POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) and various kinds of ISUCA ((F)2-(imidazol-1-yl)succinic acid)-derived lipids, which can act as pH-sensitive molecules. In order to explore such systems, we used a MARTINI-derived forcefield, previously parameterized using all-atom simulation results. We calculated the average area per lipid, the second-rank order parameter and the lipid diffusion coefficient of both lipid bilayers made of pure components and mixtures of lipids in different proportions, under neutral or acidic conditions. The results show that the use of ISUCA-derived lipids disturbs the lipid bilayer structure, with the effect being particularly marked under acidic conditions. Although more-in depth studies on these systems must be carried out, these initial results are encouraging and the lipids designed in this research could be a good basis for developing new pH-sensitive liposomes.

Construction of polyelectrolyte multilayer with anti-oxidative activity via amplified growth

Herein, precursor assembly of polyanion with pH sensitive molecule, Bis[2-(4-HydroxyPhenyl)BenzImidazole] (BHPBI), was reported to amplify layer-by-layer (LBL) exponential growth. The promoted polyelectrolyte diffusivity was responsible for the amplified LBL deposition efficiency. The release behavior of BHPBI and the anti-oxidant stress function of the film were both evaluated. The constructed multilayer film was tested to be potential drug loading and delivery system for BHPBI.

Chitosan and HPMCAS double-coating as protective systems for alginate microparticles loaded with Ctx(Ile21)-Ha antimicrobial peptide to prevent intestinal infections

The incorrect use of conventional drugs for both prevention and control of intestinal infections has contributed to a significant spread of bacterial resistance. In this way, studies that promote their replacement are a priority. In the last decade, the use of antimicrobial peptides (AMP), especially Ctx(Ile21)-Ha AMP, has gained strength, demonstrating efficient antimicrobial activity (AA) against pathogens, including multidrug-resistant bacteria. However, gastrointestinal degradation does not allow its direct oral application. In this research, double-coating systems using alginate microparticles loaded with Ctx(Ile21)-Ha peptide were designed, and in vitro release assays simulating the gastrointestinal tract were evaluated. Also, the AA against Salmonella spp. and Escherichia coli was examined. The results showed the physicochemical stability of Ctx(Ile21)-Ha peptide in the system and its potent antimicrobial activity. In addition, the combination of HPMCAS and chitosan as a gastric protection system can be promising for peptide carriers or other low pH-sensitive molecules, adequately released in the intestine. In conclusion, the coated systems employed in this study can improve the formulation of new foods or biopharmaceutical products for specific application against intestinal pathogens in animal production or, possibly, in the near future, in human health.

PH/enzyme dual sensitive and nucleus-targeting dendrimer nanoparticles to enhance the antitumour activity of doxorubicin

Direct delivery of drugs into the nucleus is a promising nanotechnology therapy, since the nucleus is one of the most important organelles controlling cell proliferation and apoptosis. Here, we report a nucleus-targeting nanocarrier for nuclear drug delivery using a pH/enzyme dual sensitive strategy. The specific ligand PGM (PKKKRKV-GFLG-Mp), composed of nuclear localization sequence (PKKKRKV), enzyme-sensitive tetrapeptide (Gly-Phe-Leu-Gly, GFLG), and pH-sensitive molecules morpholine (Mp), was modified on poly (amidoamine) (PAMAM) by maleimide active polyethylene glycol ester (NHS-PEG-MAL) to form PAMAM-PEG-PGM. Doxorubicin (DOX) was loaded into the cavity of PAMAM to prepare DOX/PAMAM-PEG-PGM. In vitro release study suggested DOX release from DOX/PAMAM-PEG-PGM nanoparticles followed pH and enzyme-triggered manner. In vitro studies showed DOX/PAMAM-PEG-PGM nanoparticles could not only promote cell internalization through the charge switching of morpholine, but also achieve nuclear internalization by the mediation of composite formed by NLS and importin α/β receptor. Further, employing H22 tumour-bearing BALB/c mice as a model, the systemic distribution and anticancer effects of nanoparticles were studied in vivo. The results indicated the nanoparticles could preferentially accumulate in the tumour site in vivo, and the tumour inhibition rate was 88.47%. In short, the nanoparticles developed could be promising in application to nucleus-targeting therapy to enhance antitumour activity.

Lateral organization of biomimetic cell membranes in varying pH conditions

Many studies have been devoted to investigation of phase separation and formation of lipid domains, which play crucial role in many biological processes. Here we present a complex study on the formation, dynamics, and stability of the phase-separated supported lipid membranes under varying pH conditions. The size and distribution of liquid-ordered (Lo) phase domains were investigated in a wide range (1.7–9.0) of buffer pH values and a strong correlation was found between the size of the Lo domains and pH of the buffer hydrating the lipid bilayer. Interestingly, the dynamics of lipids composing both Lo and Ld phase are insensitive to the pH of the buffer. Our findings demonstrate that by varying pH of the environment one can induce formation of domains with a specific size and shape without any external modification of the solid support or altering the membrane composition. Finally, we show that the architecture of the lipid membrane is stable even upon replacement of the aqueous medium with the buffer of neutral pH. Consequently, this method of patterning of Lo phase domains in biomimetic membranes is applicable to the studies involving binding of proteins or incorporation of other pH-sensitive molecules.

Carbon dots as multifunctional platform for intracellular pH sensing and bioimaging. In vitro and in vivo studies

In the present work we have obtained carbon dots (CDs) from glutathione and modified their surface with both β-Cyclodextrin and methyl-3-(4-(2-(5-((methylsulfonyl)oxy)pentyl)- 1,3-dioxo -2,3-dihydro- 1H-benzo[de]isoquinolin -6-yl)piperazin-1-yl)propanoate (FP), a pH-sensitive molecule. β-Cyclodextrin acted as a glucose source, which facilitated the entrance of the CDs within the cells and, eventually, can be used as drug-carrier to deploy bioactive compounds in the cytoplasm. FP fluorescence varied according to the pH of the medium and can be used to distinguish cancer cells due to the Warburg effect.Cytotoxicity studies revealed that, contrarily to simple CDs, carbon nanodots modified with β-cyclodextrins had an impact in cell cycle phase distribution, with no alterations of cell morphology. Likewise, CDs modified with β-cyclodextrin and FP can be used to distinguish cancer cells from non-cancerous ones due to an increase in fluorescent emission. Furthermore, this modified CDs can also be used to obtain high contrast in vitro images as well as in vivo images. These results demonstrate the potential applications of the synthetized nanoparticles both in bioimaging and as theranostic sensor and actuator.

Mesenchymal Stem Cell Regulation of Macrophage Phagocytosis; Quantitation and Imaging.

Mesenchymal stem cells (MSC) have traditionally been studied for their regenerative properties, but more recently, their immunoregulatory characteristics have been at the forefront. They interact with and regulate immune cell activity. The focus of this study is the MSC regulation of macrophage phagocytic activity. Macrophage (MΦ) phagocytosis is an important part of the innate immune system response to infection, and the mechanisms through which MSC modulate this response are under active investigation. Presented here is a method to study MΦ phagocytosis of non-opsonized zymosan particles conjugated to a pH-sensitive fluorescent molecule while in co-culture with MSC. As phagocytic activity increases and the labeled zymosan particles are enclosed within the acidic environment of the phagolysosome, the fluorescence intensity of the pH-sensitive molecule increases. With the appropriate excitation and emission wavelengths, phagocytic activity is measured using a fluorescent spectrophotometer and kinetic data is presented as changes in relative fluorescent units over a 70 min period. To support this quantitative data, the change in the phagocytic activity is visualized using dynamic imaging. Results using this method demonstrate that when in co-culture, MSC enhance MΦ phagocytosis of non-opsonized zymosan of both naive and IFN-γ treated MΦ. These data add to the current knowledge of MSC regulation of the innate immune system. This method can be applied in future investigations to fully delineate the underlying cellular and molecular mechanisms.

Mesenchymal Stem Cell Regulation of Macrophage Phagocytosis; Quantitation and Imaging

Presented here is a protocol to quantify and produce dynamic images of mesenchymal stem cell (MSC) mediated regulation of macrophage (MΦ) phagocytosis of non-opsonized yeast (zymosan) particles that are conjugated to a pH-sensitive fluorescent molecule.

A pH-sensitive ESIPT molecule with aggregation-induced emission and tunable solid-state fluorescence multicolor for anti-counterfeiting and food freshness detection

Achieving stimulus-responsive multicolor solid-state photoluminescence based on a single fluorescence compound has potential applications in anti-counterfeiting labels and visual detection. However, few such organic chromophores have these features and been used in this fields. In this study, multiple pH-sensitive sites were incorporated into a single molecule of succinimide substituted 2-(2′-hydroxyphenyl)Benzothiazole derivative (BTSA) that was Aggregation induced emission (AIE) and Excited state intramolecular proton transfer (ESIPT) active, exhibiting multilevel and highly secured anti-counterfeiting abilities. Under ammonia atmosphere, BTSA exhibits reversible switching between the green and blue fluorescence. By rationally manipulating two stimuli (acid and base), BTSA labels incredibly shows four different emission colors. The multicolor fluorescence changes are irreversible and evidently depend on the order of acid-base added. Together with smartphone that can quickly and accurately identify the multiple colors, BTSA affords a new covert fluorescent anti-counterfeiting technique with high security through the combination of reversible and irreversible fluorescence multicolor changes, as well as order and stimuli-dependent luminescent patterns. In addition, BTSA labels are successfully applied for real-time and visual monitoring the food freshness of pork and shrimp. The molecular design rationale presented here provide a new design strategy for the purposes of multicolor anti-counterfeiting and food freshness detection.

Charge-switchable, anti-oxidative molecule tuned polyelectrolyte multilayered films: Amplified polyelectrolyte diffusivity and accelerated diabetes wound healing

The local oxidizing microenvironment and bacterial infection, as crucial internal and external factors that hinder the healing of diabetic wounds, are two main issues to be addressed in the design of novel wound dressing materials. As an attempt to provide optimal solution to the aforementioned problems, free-standing (FS) polyelectrolyte multilayer films (PEMs) loaded with bis[2-(4-hydroxyphenyl) benzimidazole] (BHPB) were constructed by layer-by-layer (LBL) self-assembly method. The exponential growth of LBL deposition as well as the film degradation were extensively studied, our results revealed that by pre-assembling polycation with the pH-sensitive BHPB molecule, the polycation diffusivity was largely accelerated thus greatly amplified LBL film growth, ensuring the facile detachment of the PEMs from the substrates to achieve FS films. The increase in BHPB content leaded to faster film growth and slower film degradation and more sustainable BHPB release. In terms of multifunctionality, which was ideal for the design of wound dressing material, the obtained FS films exhibited effective antimicrobial and antioxidant properties thanks to the addition of bioactive BHPB. Notably, unlike traditional antioxidant materials that only neutralize reactive oxygen species (ROS), the BHPB-loaded films stimulated the Kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) axis, which is a vital intercellular ROS scavenging system blocked by hyperglycemia. Moreover, the wound treated by BHPB-incorporated PEMs exhibited faster healing with increased collagen deposition, enhanced angiogenesis, and rapid re-epithelialization in rat diabetic cutaneous defect models. Overall, in the perspective of cost-effectiveness and simple fabrication, the composite BHPB incorporated FS films reported in the current study showed rather promising potential in the application of diabetic wound healing.

Branched Au Nanoparticles on Nanofibers for Surface-Enhanced Raman Scattering Sensing of Intracellular pH and Extracellular pH Gradients.

The development of plasmonic-active nanosensors for surface-enhanced Raman scattering (SERS) sensing is important for gaining knowledge on intracellular and extracellular chemical processes, hypoxia detection, and label-free detection of neurotransmitters and metabolites, among other applications in cell biology. The fabrication of SERS nanosensors for optophysiology measurements using substrates such as nanofibers with a uniform distribution of plasmonic nanoparticles (NPs) remains a critical hurdle. We report here on a strategy using block copolymer brush-layer templating and ligand exchange for fabricating highly reproducible and stable SERS-active nanofibers with tip diameters down to 60 nm and covered with well-dispersed and uniformly distributed branched AuNPs, which have intrinsic hotspots favoring inherently high plasmonic sensitivity. Among the SERS sensors investigated, those with Au nanostars with short branches [AuNS(S)s] exhibit the greatest SERS sensitivity, as verified also by COMSOL Multiphysics simulations. Functionalization of the AuNS(S)s with the pH-sensitive molecule, 4-mercaptobenzoic acid, led to SERS nanosensors capable of quantifying pH over a linear range of 6.5-9.5, covering the physiological range. These pH nanosensors were shown to be able to detect the intracellular pH as well as extracellular pH gradients of in vitro breast cancer cells with minimal invasiveness and improved SERS sensitivity, along with a high spatial resolution capability.

PH-sensitive small molecule nanodrug self-assembled from amphiphilic vitamin B6-E analogue conjugate for targeted synergistic cancer therapy

To promote the targeted cancer therapy, the pH-sensitive small molecule nanodrug self-assembled from amphiphilic vitamin B6-E analogue conjugate was successfully constructed. Herein, water-soluble vitamin B6 with pKa (5.6) was chemically conjugated to lipid-soluble vitamin E succinate (α-TOS), which showed selective cancer cell killing ability and this amphiphilic small molecule vitamin conjugate could self-assemble to be free nanoparticles (NPs) and doxorubicin-loaded NPs (α-TOS-B6-NPs-DOX). The small molecule nanodrugs could perform the following characteristic: (i) stability in the sodium dodecyl sulfonate (SDS) solution and long-term storage stability in PBS via surface negative charge; (ii) tumor accumulation by enhanced penetration and retention (EPR) effect; (iii) improved cellular internalization by means of vitamin B6 transporting membrane carrier (VTC); and (iv) facilitating endosomal escape and rapid drug release for synergistic toxicity to tumor cells via charge reversal and ester hydrolysis at intracellular pH and/or esterase. Moreover, α-TOS-B6-NPs-DOX exhibited long blood circulation stability and significant tumor accumulation and inhibition with the decreased side effects in vivo. Thus, the pH-sensitive small molecule nanodrug self-assembled from amphiphilic vitamin B6-E analogue conjugate could be the potential drug carriers in targeted synergistic cancer therapy.

Influence of surfactant chain length, counterion and OrMoSil precursors on reversibility and working interval of pH colorimetric sensors

An in-depth study was conducted to determine the influence of some important parameters to produce a pH colorimetric sensor characterized by long-term stability, lack of leaching, high reversibility, repeatability and extended working range. Bromothymol Blue (BB) was used as pH sensitive molecule. Its performance was monitored while tailoring the concentration of some added cationic surfactants, together with the organic fraction of the sol-gel composition to avoid leaching phenomena, low homogeneity and to increase reversibility. Five cationic surfactants with different linear alkyl chain length (from C8 to C18) were added to a sol-gel containing BB. The C16 was chosen as optimal. The effect of the surfactant counterion was also investigated. The C16 surfactant had a pH working interval of 3 pH unit by using chloride, 1.5 with bromide and 1 pH unit with p-toluenesulfonate as counterions, respectively. Chloride was characterized by a larger inhomogeneity while p-toluenesulfonate gave homogeneity and high repeatability. The hexadecyltrimethylammonium p-toluenesulfonate was therefore chosen as the best surfactant. Leaching in alkaline medium was avoided by using the dodecyl moiety among three organic groups linked to the OrMoSil matrix (dodecyl, phenyl and methyl). Colorimetric sensor arrays made with these specifications remained stable and reversible for months.

Triplet-Triplet Annihilation Upconversion Based Nanosensors for Fluorescence Detection of Potassium.

Typical ionophore-based nanosensors use Nile blue derived indicators called chromoionophores, which must contend with strong background absorption, autofluorescence, and scattering in biological samples that limit their usefulness. Here, we demonstrate potassium-selective nanosensors that utilize triplet-triplet annihilation upconversion to minimize potential optical interference in biological media and a pH-sensitive quencher molecule to modulate the upconversion intensity in response to changes in analyte concentration. A triplet-triplet annihilation dye pair (platinum(II) octaethylporphyrin and 9,10-diphenylanthracene) was integrated into nanosensors containing an analyte binding ligand (ionophore), charge-balancing additive, and a pH indicator quencher. The nanosensor response to potassium was shown to be reversible and stable for 3 days. In addition, the nanosensors are selective against sodium, calcium, and magnesium (selectivity coefficients in log10 units of -2.2 for calcium, -2.0 for sodium, and -2.4 for magnesium), three interfering ions found in biological samples. The lack of signal overlap between the upconversion nanosensors and GFP, a common biological fluorescent indicator, is demonstrated in confocal microscope images of sensors embedded in a bacterial biofilm.