pH Insensitivity Research Articles

Fluorogenic Rhodamine-Based Chemigenetic Biosensor for Monitoring Cellular NADPH Dynamics.

Ratiometric biosensors employing Förster Resonance Energy Transfer (FRET) enable the real-time tracking of metabolite dynamics. Here, we introduce an approach for generating a FRET-based biosensor in which changes in apparent FRET efficiency rely on the analyte-controlled fluorogenicity of a rhodamine rather than the commonly used distance change between donor-acceptor fluorophores. Our fluorogenic, rhodamine-based, chemigenetic biosensor (FOCS) relies on a synthetic, protein-tethered FRET probe, in which the rhodamine acting as the FRET acceptor switches in an analyte-dependent manner from a dark to a fluorescent state. This allows ratiometric sensing of the analyte concentration. We use this approach to generate a chemigenetic biosensor for nicotinamide adenine dinucleotide phosphate (NADPH). FOCS-NADPH exhibits a rapid and reversible response toward NAPDH with a good dynamic range, selectivity, and pH insensitivity. FOCS-NADPH allows real-time monitoring of cytosolic NADPH fluctuations in live cells during oxidative stress or after drug exposure. We furthermore used FOCS-NADPH to investigate NADPH homeostasis regulation through the pentose phosphate pathway of glucose metabolism. FOCS-NADPH is a powerful tool for studying NADPH metabolism and serves as a blueprint for the development of future fluorescent biosensors.

Rational design fluorescent dyes based on morpholine derivatives for specific imaging of lysosomes and lipid droplets in living cells

Lysosomes and lipid droplets (LDs), as two crucial sub-organelles within cells, play a pivotal role in maintaining the normal cellular functions. However, rational design and synthesis of fluorophores that selectively target specific organelles still pose a challenge. Herein, we developed a series of eight fluorescent dyes (LCA1-LCA4 and LCN1-LCN4) containing morpholine moiety, which effectively targeting lysosome and LDs within cells through rational regulation of substituents in the molecular structure. Among these compounds, compounds LCA1/LCN1 containing triphenylamine group and LCA3/LCN4 containing coumarin group exhibit specific targeting towards the LDs, whereas compound LCN2 containing carbazole group demonstrates targeted localization to lysosome. Moreover, these five compounds displayed the advantages of high cellular viability, excellent resistance to interference, pH insensitivity and the ability to enter cells without washing. Compounds LCN2 and LCA1 are capable of real-time monitoring the dynamic behavior of lysosome and LDs respectively, as well as the mitophagy (LCN2) or the number and diameter variations of LDs (LCA1). The results provide a potential candidate for specific lysosomes and LDs imaging.

Surface Functionalised Optical Fibre for Detection of Hydrogen Sulphide.

Dysregulated production of hydrogen sulphide in the human body has been associated with various diseases including cancer, underlining the importance of accurate detection of this molecule. Here, we report the detection of hydrogen sulphide using fluorescence-emission enhancement of two 1,8-naphthalimide fluorescent probes with an azide moiety in position 4. One probe, serving as a control, featured a methoxyethyl moiety through the imide to evaluate its effectiveness for hydrogen sulphide detection, while the other probe was modified with (3-aminopropyl)triethoxysilane (APTES) to enable direct covalent attachment to an optical fibre tip. We coated the optical fibre tip relatively homogeneously with the APTES-azide fluorophore, as confirmed via x-ray photoelectron spectroscopy (XPS). The absorption and fluorescence responses of the control fluorophore free in PBS were analysed using UV-Vis and fluorescence spectrophotometry, while the fluorescence emission of the APTES-azide fluorophore-coated optical fibres was examined using a simple, low-cost optical fibre-based setup. Both fluorescent probes exhibited a significant increase (more than double the initial value) in fluorescence emission upon the addition of HS- when excited with 405 nm. However, the fluorescence enhancement of the coated optical fibres demonstrated a much faster response time of 2 min (time for the fluorescence intensity to reach 90% of its maximum value) compared to the control fluorophore in solution (30 min). Additionally, the temporal evolution of fluorescence intensity of the fluorophore coated on the optical fibre was studied at two pH values (7.4 and 6.4), demonstrating a reasonable overlap and confirming the compound pH insensitivity within this range. The promising results from this study indicate the potential for developing an optical fibre-based sensing system for HS- detection using the synthesised fluorophore, which could have significant applications in health monitoring and disease detection.

Ferrous ion enhanced Fenton-like degradation of emerging contaminants by sulfidated nanosized zero-valent iron with pH insensitivity

In this study, the performance and mechanism of the integrated sulfidated nanosized zero-valent iron and ferrous ions (S-nZVI/Fe2+) system for oxygen activation to remove emerging contaminants (ECs) were comprehensively explored. The S-nZVI/Fe2+ system exhibited a 2.4–8.2 times of increase in the pseudo-first order kinetic rate constant for the oxidative degradation of various ECs compared to the S-nZVI system under aerobic conditions, whereas negligible removal was observed in both nZVI and nZVI/Fe2+ systems. Moreover, remarkable EC mineralization efficiency and benign detoxification capacity were also demonstrated in the S-nZVI/Fe2+ system. We revealed that dosing Fe2+ promoted the corrosion of S-nZVI by maintaining an acidic solution pH, which was conducive to O2 activation by dissolved Fe2+ and surface-absorbed Fe(II) to produce •OH. Furthermore, the generation of H* was enhanced for the further reduction of Fe(III) and H2O2 to Fe(II) and •O2–, resulting in the improvement of consecutive single-electron O2 activation for •OH production. Additionally, bisphenol A (BPA) degradation by S-nZVI/Fe2+ was positively correlated with the S-nZVI dosage, with an optimum S/Fe molar ratio of 0.15. The Fenton-like degradation process by S-nZVI/Fe2+ was pH-insensitive, indicating its robust performance over a wide pH range. This study provides valuable insights for the practical implementation of nZVI-based technology in achieving high-efficiency removal of ECs from water.

Selective visualization of nitric oxide in living cells and tissue slices using a mitochondria-targetable fluorescent probe

Nitric oxide (NO) is an important messaging molecule which plays a vital role in many biological systems. Because the endogenous NO is synthesized mainly in the mitochondria of mammalian cells, the detection of NO in mitochondria is of great importance for studying the biological functions of NO. Here, a novel fluorescence turn-on probe (MTNO) for monitoring the mitochondrial NO is designed and synthesized. The fluorescence intensity of MTNO exhibits a ∼430-fold enhancement in response to NO with a detection limit as low as 14 nM. MTNO also shows good mitochondrial localization, pH insensitivity and high selectivity to NO. With the aid of MTNO, the imaging of exogenous and endogenous NO in living cells and tissue slices is achieved through two-photon microscopy.

Low duty cycle pulse trains for exchange rate insensitive chemical exchange saturation transfer MRI.

To introduce and validate a pulse scheme that uses low duty cycle trains of π-pulses to achieve saturation that is relatively insensitive to exchange rate yet linearly dependent on labile proton concentration. Simulations were performed to explore the exchange rate sensitivity of π-pulse trains and continuous wave chemical exchange saturation transfer (CEST) signals. Creatine phantoms with varying pH and varying concentrations were imaged to demonstrate pH insensitivity and concentration dependence of low duty cycle π-pulse saturation. Simulations show decreasing the duty cycle of π-pulse saturation decreases peak sensitivity to exchange rate, and this range of insensitivity can be tuned to different exchange rates through average B1 power. The range of insensitivity is unaffected by changes in relaxation and magnetization transfer, while the sensitivity of CEST signal maintains linear dependence on labile proton concentration. Under B1,avg = 0.48 μT, 30mM creatine with pHs ranging between 6.36 and 8.21 exhibited CEST contrast ranging between ~6 and 11% under continuous wave and ~4% across all pHs using 10% duty cycle π-pulses. Imaging these phantoms using duty cycles of 5, 10, 25, and 50% showed decreasing pH sensitivity with decreased duty cycle. Creatine phantoms with varied concentrations and pHs reveal that π-pulse train saturation exhibited stricter correlation to concentration at lower DCs. Low DC π-pulse train is an easy-to-implement way of providing labile proton concentration-dependent CEST MRI signal that is insensitive to exchange rate. This approach can be useful in studies where a change of chemical exchange rate may interfere with accurate assessments of physiology or pathology.

Proximity-Dependent Switchable ATP Aptasensors Utilizing a High-Performance FRET Reporter.

Here, fluorescent molecular rotors are employed to develop a new type of high-performance FRET system with large Stokes shift, high photostability, and pH insensitivity, showing great promise for use in proximity-dependent DNA aptasensors. Two carboxylated benzothiazole-based molecular rotors are synthesized, displaying bright green and red fluorescence once labeled to DNA. In the proximity state, an efficient FRET occurs between the two dyes, comparable to that of the most commonly used Cy3/Cy5 pair. Similar phenomena are also observed if naphthothiazole-based analogues are adopted. Our developed FRET pair is then attached to the two parts of a split ATP aptamer in a dimeric DNA nanoscaffold controlled by a bimolecular i-motif. In this way, a pH-switched proximity-induced fluorescent ATP aptasensor is constructed, with good sensitivity, selectivity, and reconfiguration. Furthermore, by altering the linker length of the switching unit, the proximity effect is investigated systematically, providing new insight into DNA proximity reactions and their roles in some physiological processes.

Molecular Scylla and Charybdis: Maneuvering between pH Sensitivity and Excited-State Localization in Ruthenium Bi(benz)imidazole Complexes.

Bi(benz)imidazoles (b(b)im) acting as N,N-chelates in ruthenium complexes represent a unique class of ligands. They do not harbor metal-to-ligand charge-transfer (MLCT) excited states in ruthenium polypyridyl complexes upon visible-light excitation provided that no substitution is introduced at the N atoms. Hence, they can be used to steer light-driven electron-transfer pathways in a desired direction. Nonetheless, the free N atoms are susceptible to protonation and, hence, introduce highly pH-dependent properties into the complexes. Previous results for ruthenium complexes containing R2bbim ligands with alkylic or arylic N,N'-substitution indicated that, although pH insensitivity was accomplished, unexpected losses of spectator ligand features incurred simultaneously. Here, we report the synthesis and photophysical characterization of a series of differently N,N'-alkylated b(b)im ligands along with their corresponding [(tbbpy)2Ru(R2b(b)im)](PF6)2 complexes (tbbpy = 4,4'-tert-butyl-2,2'-bipyridine). The data reveal that elongation of a rigid ethylene bridge by just one methylene group drastically increases the emission quantum yield, emission lifetime, and photostability of the resultant complexes. Quantum-chemical calculations support these findings and allow us to rationalize the observed effects based on the energetic positions of the respective excited states. We suggest that N,N'-propylene-protected 1H,1'H-2,2'-biimidazole (prbim) is a suitable spectator ligand because it stabilizes sufficiently long-lived MLCT excited states exclusively localized at auxiliary bipyridine ligands. This ligand represents, therefore, a vital building block for next-generation photochemical molecular devices in artificial photosynthesis.

Dimethylamino naphthalene-based fluorescent probes for hydrogen sulfide detection and living cell imaging.

Hydrogen sulfide shows great importance in various physiological and biochemical processes. The development of fluorescence probes for facile and efficient detection of H2S has attracted increasing attention of researchers. Herein, we synthesized two fluorescence probes based on simple naphthalene structure for detection of H2S. Upon reaction with H2S, the probe DN-DM exhibited a red fluorescence emission with large Stokes shift. The probe showed high sensitivity, pH insensitivity and good selectivity for H2S over other analytes including common biothiols. The detection mechanism was based on the thiolysis of the dinitrophenyl ether moiety, which was confirmed by 1H NMR spectral analysis. The DFT calculation was also performed for a deeper understanding of the photophysical properties. In addition, these probes showed good cell-membrane permeability and could be utilized for detection of H2S in living cells.

An innovative pH-independent magnetically separable hydrogel for the removal of Cu(II) and Ni(II) ions from electroplating wastewater

A novel magnetic anionic hydrogel (nFeMAH), synthesized via a facile method, was characterized by XRD, VSM, SEM, TEM, FTIR, XPS, and ζ-potential measurement. Over the tested range of pH 2 to 12, the surface of the nFeMAH was permanently negative with a ζ-potential of −35 to −45 mV. The adsorption kinetics and capacity of nFeMAH were studied. Within 60 min, equilibrium was achieved with maximum adsorption capacities of 102 mg Cu(II)/g and 93 mg Ni(II)/g. The pseudo-second-order kinetics model was well-matched with the experimental data, whereas the Langmuir isotherm model agreed well with the isotherm data. The magnetic separation efficiency of nFeMAH remained above 90% after 20 cycles of adsorption-desorption, whereas the Ni(II) removal efficiency dropped from 92 to about 75% after the first cycle. The magnetic separation efficiency of nFeMAH was consistently high (99%). The major mechanism of metal removal by nFeMAH was ion exchange but there also was evidence for formation of metal oxides. Therefore, the application of nFeMAH for treating electroplating wastewater can be a desirable option when considering its superior performance in the adsorptive treatment, i.e., pH insensitivity, fast adsorption kinetics, high reusability, and consistency in magnetic separation.

Cy7-Tocilizumab/Fab(Tocilizumab): Near Infrared Fluorescence In Vivo Imaging of Multiple Myeloma

Introduction:Multiple Myeloma (MM) is an incurable bone marrow cancer characterized by the proliferation of malignant plasma cells. Interleukin-6 (IL-6) is a key player molecule related to growth, survival and proliferation of MM cells. Tocilizumab is a humanized monoclonal antibody directed against the soluble and membrane IL-6 receptor. We have previously radiolabeled Tocilizumab and its Fab´s fragments with 99mTechnetium, revealing its potential role as MM radiotracers for targeting IL-6 expression in vivo [1, 2]. Near-infrared (650-900 nm) fluorescence (NIRF) cyanine (Cy) dye has been used for labeling several molecules for optical imaging due to their properties, such as small size, good aqueous solubility, and pH insensitivity between pH 3 and 10.Objectives: Label Tocilizumab and Fab´s (Tocilizumab) with Cy7 and evaluate its potential role as MM imaging agent.Methodology: IL-6R expression in MM cell lines (U266, NCI-H929 and MM1S) was confirmed by laser confocal microscopy. Tocilizumab fragmentation was carried out with papain and, once purified, fragments were identified by MALDI/TOF and SDS-PAGE. Cy7-Tocilizumab / Fab(Tocilizumab) were synthesized through nucleophilic substitution reaction between monofunctional N-hydroxysuccinimide ester (Cy7-NHS) and Tocilizumab / Fab(Tocilizumab) [3]. After purification, the conjugates were characterized by spectrophotometry. For in vivo imaging, Cy7-Tocilizumab/Fab(Tocilizumab) (1 nmol) were injected intravenously in MM1S tumor-bearing Balb/c nude mice and were imaged with near-infrared fluorescence (NIRF) after 0, 1, 2, 6 and 24 h post-injection of Cy7-Tocilizumab and after 0.5, 2, 6, 24, 48 and 72 h post-injection of Cy7-Fab(Tocilizumab).Results: Laser confocal microscopy showed that MM cell lines express high levels of IL-6R. Pure and homogeneous Fab-Tocilizumab fragments were produced. Tocilizumab and Fab (Tocilizumab) were successfully labeled with Cy7 as shown by spectrophotometry. Non-invasive NIRF in vivo imaging of MM tumor-bearing Balb/c nude mice allowed us to distinguish tumors up to 24 h and 72 h post-injection of Cy7-Tocilizumab and Cy7-Fab(Tocilizumab), respectively (Figures A and B).Conclusions: MM cell lines express IL-6R. Cy7 labeled Tocilizumab/Fab(Tocilizumab) has the potential to become optical imaging agents for IL-6R expressing tumors such as MM, being useful to guiding surgical excision of tumors and biopsies, that merits further evaluation.Acknowledgments: Agencia Nacional de Innovación e Investigación - Uruguay (ANII), Roche Laboratories, Pro.In.Bio (Uruguay), PEDECIBA Química (Uruguay)) and Comisión Sectorial de Investigación Científica-Universidad de la República-Uruguay (CSIC, UdelaR) I+D Grupos Oncología Nuclear.Disclosures: No relevant conflicts of interest to declare.

South American characids share very similar ionoregulatory characteristics

We examined ionoregulatory characteristics of four Characids from diverse locations in South America, emperor tetras (Nematobrycon palmeri), penguin tetras (Thayeria boehlkei), serpae tetras (Hyphessobrycon eques), and rosy tetras (Hyphessobrycon rosaceous). When held in 100 μmol L−1 Na+ water, tetras had Jmax values over 1100 nmol g−1 h−1, and Km values below 60 μmol L−1. When held in 1 mmol L−1 Na+ water kinetic parameters were unchanged. Low pH had no effect on Na+ uptake (JinNa). At pH 3.25, Na+ loss (JoutNa) was stimulated 35–85% in two of the four species. To test the linkage of JinNa to NH3 and H+ extrusion we measured JinNa during exposure to 1 mmol L−1 NH4Cl (HEA) and 100 μmol L−1 Acetazolemide (AZ). HEA stimulated JinNa of emperor tetras by 40%, but inhibited JinNa of penguin tetras by 50%; the two remaining species were unaffected. AZ (an inhibitor of carbonic anhydrase) inhibited JinNa of serpae tetras by 40%, but had no effect on the others. All tetras displayed ionoregulatory characteristics that are very similar to each other, which supports the argument that these physiological traits may be ancestral for this group and pre-date colonization of the Rio Negro. The novel finding that, Jmax and Km did not change after acclimation to 1 mM Na+ water indicates that, unlike in other species examined uptake is not plastic. The HEA and AZ results, along with pH insensitivity suggest Na+ uptake is not coupled to H+ extrusion or NH3 excretion and leaves the exact mechanism involved unclear.

A biomolecule-based fluorescence chemosensor for sequential detection of Ag+ and H2S in 100% aqueous solution and living cells

The silver ions (Ag+) and hydrogen sulfide (H2S) play extremely an important role in the organism and the environment, while there lack a novel probe with biodegradability, good water solubility, and good biocompatibility to detect Ag+ and H2S individually. Herein, a novel fluorescence chemosensor DP (Dansyl-Leu-Leu-Cys-Acp-Asp-OH) based on biomolecular peptide has been designed and synthesized, which can be used in the detection of intracellular and environmental Ag+ and H2S. The DP exhibited high selectivity and sensitivity, excellent cell permeation, biodegradability, good water solubility, and pH insensitivity over a biologically relevant range, with detection limits of 61.7 nM and 74.0 nM for Ag+ and H2S, respectively. This work can inspire the design of multi-functional fluorescence sensor based on beacon peptides by modifying lateral and terminal groups for imaging applications in physiological and pathological events.

TnI Structural Interface with the N-Terminal Lobe of TnC as a Determinant of Cardiac Contractility

The heterotrimeric cardiac troponin complex is a key regulator of contraction and plays an essential role in conferring Ca2+ sensitivity to the sarcomere. During ischemic injury, rapidly accumulating protons acidify the myoplasm, resulting in markedly reduced Ca2+ sensitivity of the sarcomere. Unlike the adult heart, sarcomeric Ca2+ sensitivity in fetal cardiac tissue is comparatively pH insensitive. Replacement of the adult cardiac troponin I (cTnI) isoform with the fetal troponin I (ssTnI) isoform renders adult cardiac contractile machinery relatively insensitive to acidification. Alignment and functional studies have determined histidine 132 of ssTnI to be the predominant source of this pH insensitivity. Substitution of histidine at the cognate position 164 in cTnI confers the same pH insensitivity to adult cardiac myocytes. An alanine at position 164 of cTnI is conserved in all mammals, with the exception of the platypus, which expresses a proline. Prolines are biophysically unique because of their innate conformational rigidity and helix-disrupting function. To provide deeper structure-function insight into the role of the TnC-TnI interface in determining contractility, we employed a live-cell approach alongside molecular dynamics simulations to ascertain the chemo-mechanical implications of the disrupted helix 4 of cTnI where position 164 exists. This important motif belongs to the critical switch region of cTnI. Substitution of a proline at position 164 of cTnI in adult rat cardiac myocytes causes increased contractility independent of alterations in the Ca2+ transient. Free-energy perturbation calculations of cTnC-Ca2+ binding indicate no difference in cTnC-Ca2+ affinity. Rather, we propose the enhanced contractility is derived from new salt bridge interactions between cTnI helix 4 and cTnC helix A, which are critical in determining pH sensitivity and contractility. Molecular dynamics simulations demonstrate that cTnI A164P structurally phenocopies ssTnI under baseline but not acidotic conditions. These findings highlight the evolutionarily directed role of the TnI-cTnC interface in determining cardiac contractility.

Nitrate Ion Selective Electrode Based on Ion Imprinted Poly(N-methylpyrrole)

A poly(N-methylpyrrole) based ion selective electrode (ISE) has been prepared by electro-polymerization of N-methylpyrrole using potassium nitrate as the supporting electrolyte. Electrochemical and chemical variables were used to optimize the potentiometric response of the electrodes and to maximize the selectivity for nitrate over potential interferences. The selectivity, longevity and stability of the ion-imprinted polymer give this electrode advantages over traditional nitrate ISEs. The best prototype electrode exhibits a linear potential response to nitrate ion within the concentration range of 5.0 × 10−6 to 0.1 M nitrate with a near Nernstian slope of −56.3 mV per decade (R2 = 0.9998) and a strong preference for the nitrate ion over other anions. The selectivity coefficients of the electrode were evaluated by the fixed interference method. The use of N-methylpyrrole has advantages over pyrrole in terms of selectivity and pH insensitivity.

Hagfish slime exudate stabilization and its effect on slime formation and functionality.

ABSTRACTHagfish produce vast amounts of slime when under attack. The slime is the most dilute hydrogel known to date, and is a highly interesting material for biomaterial research. It forms from a glandular secrete, called exudate, which deploys upon contact with seawater. To study slime formation ex vivo and to characterize its material properties, stabilization of the sensitive slime exudate is crucial. In this study, we compared the two main stabilization methods, dispersion in high osmolarity citrate/PIPES (CP) buffer and immersion in oil, and tested the influence of time, temperature and pH on the stability of the exudate and functionality of the slime. Using water retention measurements to assess slime functionality, we found that CP buffer and oil preserved the exudate within the first 5 hours without loss of functionality. For longer storage times, slime functionality decreased for both stabilization methods, for which the breakdown mechanisms differed. Stabilization in oil likely favored temperature-sensitive osmotic-driven swelling and rupture of the mucin vesicles, causing the exudate to gel and clump. Extended storage in CP buffer resulted in an inhibited unraveling of skeins. We suggest that a water soluble protein glue, which mediates skein unraveling in functional skeins, denatures and gradually becomes insoluble during storage in CP buffer. The breakdown was accentuated when the pH of the CP buffer was raised from pH 6.7 to pH 8.5, probably caused by increased denaturation of the protein glue or by inferior vesicle stabilization. However, when fresh exudate was mixed into seawater or phosphate buffer at pH 6-9, slime functionality was not affected, showing pH insensitivity of the slime formation around a neutral pH. These insights on hagfish exudate stabilization mechanisms will support hagfish slime research at a fundamental level, and contribute to resolve the complex mechanisms of skein unraveling and slime formation.

Open-Chain Crown-Ether-Derived Two-Photon Fluorescence Probe for Real-Time Dynamic Biopsy of Mercury Ions

A novel two-photon fluorescence probe for Hg2+ derived from bis(styryl)terephthalonitrile, as a two-photon fluorophore, and bis[2-(2-hydroxyethyl sulfanyl) ethyl]amino group (ionophore), as a novel Hg2+ ligand, was developed. The probe possesses small molecule size, large two-photon absorption cross-section (1067 GM) in H2O, non-cytotoxic effect, long wavelength emission at 588 nm, large Stokes shift (121 nm), excellent photostability, high water solubility, good cell permeability, and pH insensitivity in the biologically relevant range. The probe can selectively detect Hg2+ ions in live cells and living tissues without interference from other metal ions and the membrane-bound probes, and its quenching constant is 8.73 × 105 M–1.

A novel 1,8-naphthalimide derivative as an efficient silver(I) fluorescent sensor

Novel N-n-butyl-1,8-naphthalimide derivatives with mono- or di-substitution to the C-4 position containing a N atom and a carbonyl have been synthesized as a efficient silver(I) fluorescent sensor. The ligand structures were confirmed by IR, NMR, MS, and elemental analysis. A highly sensitive spectrofluorometric method based on chelation-assisted fluorescence quenching was developed for trace-level detection, estimation and speciation studies of silver(I) in ethanol-water. The ligand exhibits emission and excitation maxima at 525nm and 420nm, respectively. The novel fluorescent sensor exhibited selective binding toward Ag(I) and pH insensitivity over a biologically relevant range. The interference of foreign ions was observed to be negligible.

A BODIPY‐Based Water‐Soluble Fluorescent Probe for Mitochondria Targeting

A water‐soluble fluorescent mitochondrial targeting probe is reported. The probe bears a BODIPY‐based fluorophore, two triphenylphosphonium (TPP) groups as mitochondrial targeting moieties, and two PEG groups to increase water solubility and enhance biocompatibility. As a mitochondria‐selective fluorescent probe, the new BODIPY derivative exhibited many desirable properties, such as water solubility, high fluorescence quantum yield, solvent independent photophysical properties, pH insensitivity, good photostability, low cytotoxicity, and excellent mitochondrial targeting ability. Fluorescence microscopy colocalization experiments demonstrated the ability of this new probe to target mitochondria within living cells for bioimaging.

Two-Photon Probes for Lysosomes and Mitochondria: Simultaneous Detection of Lysosomes and Mitochondria in Live Tissues by Dual-Color Two-Photon Microscopy Imaging.

Novel two-photon (TP) probes were developed for lysosomes (PLT-yellow) and mitochondria (BMT-blue and PMT-yellow). These probes emitted strong TP-excited fluorescence in cells at widely separated wavelength regions and displayed high organelle selectivity, good cell permeability, low cytotoxicity, and pH insensitivity. The BMT-blue and PLT-yellow probes could be utilized to detect lysosomes and mitochondria simultaneously in live tissues by using dual-color two-photon microscopy, with minimum interference from each other.