A Supersmall Single-Cell Nanosensor for Intracellular K + Detection
2020; Chinese Chemical Society; Volume: 3; Issue: 9 Linguagem: Inglês
10.31635/ccschem.020.202000451
ISSN2096-5745
AutoresXiaomei Shi, Yi‐Fan Ruan, Haiyan Wang, Weiwei Zhao, Jing‐Juan Xu, Hong‐Yuan Chen,
Tópico(s)Electrochemical Analysis and Applications
ResumoOpen AccessCCS ChemistryRESEARCH ARTICLE1 Sep 2021A Supersmall Single-Cell Nanosensor for Intracellular K+ Detection Xiao-Mei Shi, Yi-Fan Ruan, Hai-Yan Wang, Wei-Wei Zhao, Jing-Juan Xu and Hong-Yuan Chen Xiao-Mei Shi State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023 , Yi-Fan Ruan State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023 , Hai-Yan Wang State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023 , Wei-Wei Zhao *Corresponding authors: E-mail Address: [email protected] E-mail Address: [email protected] State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023 , Jing-Juan Xu State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023 and Hong-Yuan Chen *Corresponding authors: E-mail Address: [email protected] E-mail Address: [email protected] State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023 https://doi.org/10.31635/ccschem.020.202000451 SectionsSupplemental MaterialAboutAbstractPDF ToolsAdd to favoritesDownload CitationsTrack Citations ShareFacebookTwitterLinked InEmail Intracellular potassium ions (K+) play pivotal roles in many physiological processes. Several K+ sensors have been developed for probing cellular K+ fluctuations. Nevertheless, the existing solutions are incompatible and impractical for intracellular K+ probing. Herein, we report a supersmall biomimetic K+ nanosensor to serve as a transmembrane vector capable of electrochemically detecting intracellular K+ in a minimally invasive manner. The sensitive and reversible response of this nanosensor stems from the synergy between a supersmall nanopipette with an orifice of ca. 20 nm and the K+-dependent structural change of the G-quadruplex DNA decorated within the nanopipette. This nanosensor is demonstrated to be an accessible tool for tracking stimulus-induced K+ fluctuation under physiological conditions. Download figure Download PowerPoint Introduction Healthy cells tightly manage their cellular ionic compositions, concentrations, and distributions, which are critical to maintain cellular physiology.1 Upset to the normal intracellular ion homeostasis and dynamics are frequently associated with severe dysfunctions and pathologies, such as metabolic disorders, neurotoxicity, and malignant tumors.2,3 Significantly, potassium ions (K+), as the fundamental metal cation with the highest concentration in the cytoplasm, play important roles in biological activities.4–6 The normal intracellular concentration of K+ is approximately 100−150 mM, whereas the extracellular value is as low as 3.5−5 mM. Disturbances in such a homeostasis are usually involved with pathological alterations. For instance, acute imbalance of intracellular K+ has been recognized in the primary stage of apoptotic volume decrease (AVD),7 a ubiquitous characteristic of programmed cell death. A high level of intracranial extracellular K+ is associated with traumatic head injuries that have been linked with a poor survival rate.8 Due to the significance of K+ in biological functions, techniques for probing cellular K+ are of vital importance in many arenas from cellular biology to biomedical diagnostics. Current clinical methods, such as ion-selective electrodes (ISEs), are capable of K+ measurements only in serum and urine.9–11 Although ion-selective liquid membranes had been front filled within glass pipettes to develop various ion-selective glass microprobes (ISGMs), they suffered from a set of limitations such as unavoidable signal attenuation and cell destructiveness during intracellular insertion due to their large sizes.12–17 Recently, many artificial K+-selective nanopores created in the films18,19 exhibited high sensitivity toward K+, but these electrochemical20 or fluorescent21 techniques were in principle limited for transmembrane application. Very recently, two optical methods have been proposed for intracellular K+ measurements,22,23 which necessitated laborious synthesis and delivery of nonnative fluorescent indicators into the cytosol. Possible physiological perturbations and light-induced toxicity may occur within the cells due to the generation of singlet oxygen and other free radicals. Additionally, possible photobleaching of fluorescent indicators would be unfavorable to the stable and long-term detection. Obviously, to advance our knowledge of K+-correlated diseases and their ultimate mechanisms, development of advanced techniques addressing intracellular K+, while keeping the cell within its natural environment, is highly needed. Experimental Section Reagents and apparatus Analytical grade chemicals were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai,China; unless stated otherwise) and used as received. The nucleotide sequences were obtained from Sangon Biotech. Co., Ltd. (Shanghai, China). The quartz capillaries and corresponding laser micropuller (P-2000) were purchased from Sutter Instruments (Novato, CA, USA). The ionic current measurements were processed using a CASCADE Summit 12000B-M with the KEITHLEY 4200-SCS. The further details of other materials, equipment, conditions, and approaches are available in the Supporting Information. Computational methods The finite element method (FEM) was used to study the mechanism of ion transport in nanofluidic circuits combined with the Poisson–Nernst–Planck equation. The computed model of nanocapillary was fabricated by using an axial symmetry structure. More details are available in the Supporting Information. Results and Discussion Fabrication of the supersmall intracellular K+ nanosensor Inspired by the biological K+ channel ( Supporting Information Figure S1) and using a nanopipette,24–32 here, we describe a supersmall transmembrane K+ nanosensor capable of electrochemical probing of intracellular K+. The nanosensor was engineered by functionalization of a small nanopipette with the G-quadruplex (G4) DNA sequence, which exhibited K+-dependent conformational transformation33 and [2.2.2-cryptand]-enabled recyclability34 (Figure 1a). As shown, the nanopipette was fabricated with an approximately 20 nm diameter orifice (Figure 1b and Supporting Information Figure S2), which enabled easy cell penetration while avoiding cell damage. The employed G4 DNA sequence was 5′-HOOC-(CH2)6-AAA AAA GGG TTA GGG TTA GGG TTA GGG-3′, thus, the height of the formed G4 DNA was estimated as 3 nm by inferring that a three-base-pair codon was ca. 1 nm. The K+-dependent structural change of the G4 DNA sequence was confirmed by circular dichroism (CD) spectroscopy (Figure 1c). As the K+ concentration increased, the gradual enhancement of the positive peak at ca. 295 nm with a crossover at ca. 265 nm indicated the typical G4 DNA conformation.35–37 To develop the nanosensor, the nanopipette was subjected to sequential silanization and amidation reactions to immobilize the DNA strands ( Supporting Information Figure S3) and then tested for K+ response, which was monitored by i–v curves (Figure 1d). As shown, the pristine nanopipette exhibited a negatively charged surface (black curve), while the 3-aminopropyltriethoxysilane (APTES) modification38 induced a positively charged surface (red curve). Functionalization of the DNA sequence reversed the surface property due to the highly negatively charged DNA sequences (blue curve).20 In view of the normal intracellular K+ range, the as-developed nanosensor was then treated with 50 mM (orange curve), 100 mM (magenta curve), and 200 mM K+ (green curve), respectively. Lower ionic current responses were observed with the higher K+ concentration. From the i–t measurements, the change of the ionic current magnitudes associated with different gating states was clearly reflected (Figure 1e). The reduced ionic currents indicated that the K+-provoked packing of the G4 DNA strands could be sensibly transduced by the small 20 nm aperture. A numerical simulation was also conducted, and the computational results confirmed the experimental observation, that is, the simulated currents displayed a downward trend with reducing effective diameter ( Supporting Information Figures S4 and S5 and Table S1). Figure 1 | (a) Design of a biomimetic K+ nanosensor using a G4-DNA formation/deformation strategy. (b) Cross-sectional and top view SEM of the nanopipette with an ultrasmall 20 nm orifice. Inset below: digital picture of the nanopipettes. (c) CD spectra of G4 DNA (5′-GGG TTA GGG TTA GGG TTA GGG-3′) against variable K+ concentrations. (d) Stepwise i–v curves of the nanopipette functionalization and its responses to variable K+ concentrations. (e) Corresponding ionic currents as-obtained by i–t measurements (bias at −1.0 V), which were performed by immersing the nanosensor in 20 mM Tris–HCl (pH 7.4) containing 40 mM LiCl (to replace KCl) after exposing the nanosensor to K+ solutions for 5 s. SEM, scanning electron microscopy; CD, circular dichroism. Download figure Download PowerPoint Selectivity and reversibility of the nanosensor The selectivity of the nanosensor was studied by exposing it to a series of interference species and a mixture of all species (Figure 2a). As shown, the ionic responses from K+ and mixture were much higher than other species, indicating the signal change was caused by K+. The specificity was also confirmed by CD spectra of the employed DNA sequence upon treatments with different common metal ions (Figure 2b), which revealed that these ions could not cause conformational change of the DNA sequence. The reversibility was subsequently studied by treating the nanosensor with 50 mM K+ and 0.1 M [2.2.2]-cryptand alternately (Figure 2c). As shown, the highly reversible switching of signals demonstrated the excellent efficiency of K+ and [2.2.2-cryptand]-enabled recyclability as well as good durability for repeated uses. Similarly, the CD spectra also confirmed the higher binding constant of [2.2.2-cryptand] with K+ than the DNA sequence (Figure 2d). As shown, 100 mM K+ treatment caused the relaxed DNA (black curve) to form the G4 DNA structure (green curve). Addition of the 100 mM [2.2.2-cryptand] resulted in its deformation (dark yellow curve), whereas further addition of 100 mM K+ induced the formation of G4 DNA again (magenta curve). The reproducibility of the nanosensor was demonstrated within eight nanopipettes from the same batch ( Supporting Information Figure S6). Incidentally, control experiments using the nonspecific DNA sequences were performed, which revealed that K+ ion treatment of the nonspecific sequences did not alter the CD spectra or the electrochemical signals ( Supporting Information Figure S7). Together these results implied the potential of the nanosensor for further implementation. Figure 2 | (a) Selectivity study of the nanosensor toward different interference species, including 100 mM K+, 100 mM Na+, 5 mM Mg2+, 180 nM Zn2+, 100 μM DA, 100 μM AA, 100 μM UA, cell medium [commercial DMEM (10% fetal bovine serum, FBS)], and their mixed sample. (b) CD spectra of the used DNA sequence (5′-GGG TTA GGG TTA GGG TTA GGG-3′) upon treatment with different common metal ions, including 50 mM Li+, 15 mM Na+, 5 mM Mg2+, and 180 nM Zn2+. (c) Reversibility of the nanosensor enabled by repeated treatments with 50 mM K+ and 100 mM [2.2.2]-cryptand. (d) CD spectra of the 10 μM DNA sequence upon K+ and cryptand treatments. The buffer for preparing the interference and DNA solution was 20 mM Tris–HCl (pH 7.4), and the electrochemical signals were collected at bias of −1.0 V. DA, dopamine; AA, ascorbic acid; UA, uric acid; DMEM, Dulbecco's modified Eagle's medium; CD, circular dichroism. Download figure Download PowerPoint Nondestructive probing of intracellular K+ level The nanosensor was then applied to analysis of intracellular K+ (Figure 3a). As shown, its ultrasmall nanotip could easily penetrate the cell (Figure 3b and Supporting Information Figure S8 and Movie S1). Ionic currents corresponding to the approaching and penetrating processes were also monitored by i–t characteristics. After a relatively stable ionic response before contact (from 0 to 15 s), the penetration (at ca. 15 s) of the nanosensor against the cell membrane led to a sudden jump of the ionic current and a subsequent recovery (after ca. 15 s) (Figure 3b, inset).39 To evaluate the impact of nanosensor insertion against the cell, cellular insertion and withdrawal of the nanosensor against the MCF-7 cell was repeated six times, and it was observed that the targeted cell kept its morphology ( Supporting Information Figure S9). The corresponding membrane integrity ( Supporting Information Figure S10) was also confirmed by using the fluorescent probe propidium iodide (PI).40 In addition, the cells were stained by Hoechst 33342 to verify the cell viability after 60 min penetration (Figure 3c). The Hoechst 33342 dye41,42 has a certain membrane permeability. The living cell is slightly stained and shows low blue fluorescence, but dead cells show a brighter blue fluorescence. After 60 min insertion, the fluorescence micrograph of the cells remained unchanged, indicating the normal viability maintenance of cells even with extended penetration by the nanosensor ( Supporting Information Figure S11). These results evidenced the feasibility for probing intracellular K+ with a DNA-based nanosensor while largely preserving the cell viability. In a parallel test, the ionic responses of the intracellular K+ from four individual MCF-10A, Hela, HepG2, and MCF-7 cells were successfully recorded (Figure 3d). Figure 3 | (a) Application of the nanosensor for probing intracellular K+. (b) Cell-penetrating assisted by a microscope and a 3D micromanipulator; inset was the ionic response of the nanosensor as it's penetrating the cell membrane (bias at +1.0 V). (c) Bright-field and confocal fluorescence micrographs of Hoechst 33342 stained MCF-7 cells before penetration and after withdrawal of the nanopipette. The excitation wavelength was 352 nm. (d) The responses of the nanosensor toward the MCF-10A, Hela, HepG2, and MCF-7 cells. The i–t curves were recorded by immersing the nanosensor in 20 mM Tris–HCl (pH 7.4) containing 40 mM LiCl (bias at −1.0 V) after holding the nanotip in cytosol for 5 s. Download figure Download PowerPoint Evaluation of drug effects Due to the importance of stable intracellular K+ concentration, the potential of the nanosensor to follow the drug-stimulated K+ fluctuations was further investigated. The resveratrol (Res)-induced apoptosis43 of the MCF-7 cell was performed with the assistance of 4,6-diamidino-2-phenylindole (DAPI) staining44 (Figures 4a and 4b and Supporting Information Figure S12) and Calcein AM/PI staining ( Supporting Information Figure S13). As shown, 4 h Res treatment caused obvious loss of the cellular volume, and the chromatin condensation was proved by the DAPI fluorescence, confirming the occurrence of primary stage AVD45 when intracellular K+ decreased to 30 mM.7 The Calcein AM/PI staining experiment indicated a decreased cellular viability but a retained membrane integrity of the early apoptotic cells. Incidentally, the small quantity of the dimethyl sulfoxide (DMSO) solvent had no effect on the cell, which was also confirmed by the Calcein AM/PI observation ( Supporting Information Figures S14–S16). Then, the contrast study of a single normal cell and apoptotic cell was conducted using the nanosensor (Figures 4c and 4d). The bright imaging clearly demonstrated the volume loss of the Res-treated cell. More importantly, the distinct ionic responses (black and green curves in Figure 4e) strongly evidenced the acute decrease of intracellular K+ in the treated cell, further indicating the potency of the nanosensor in differentiating drug effects on cellular behaviors. Under identical experimental conditions, the effects of adriamycin (Adm)46,47 and baicalin (Bai)48 were then investigated (orange and blue curves in Figure 4e). Comparatively, Res-induced AVD the best among the three drugs. Based on the data obtained from the as-prepared nanosensor (Figures 1d and 1e), the average K+ concentration was estimated as ca. 124.44 mM (±10.06 mM) in MCF-7 cells, which was consistent with reported literature,45 and as ca. 35.85 (±15.06 mM), 58.85 (±17.69 mM), 87.55 (±14.84 mM) in Res-, Adm-, and Bai-induced cells, respectively. These results verified the sensitivity of the nanosensor for single-cell K+ detection and the evaluation of drug effect. The decreasing trend of the intracellular K+ concentration within 4 h after Res treatment was revealed ( Supporting Information Figure S17). Statistical studies with 50 cells per group were also conducted (Figure 4f). These studies clearly demonstrated its capability for probing intracellular K+ fluctuation down to the single-cell level. Figure 4 | Confocal fluorescence micrographs of (a) normal DAPI-stained MCF-7 cells and (b) apoptotic MCF-7 cells induced by 4 h treatment of Res. Bright-field imaging of (c) single MCF-7 cell and (d) the apoptotic cell that was penetrated by the nanosensor. (e) The corresponding ionic responses from the normal and apoptotic cell treated with Res, Adm, and Bai. (f) The corresponding statistical analysis of 50 cells per group. All the electrochemical signals were collected at bias of −1.0 V. DAPI, 4,6-diamidino-2-phenylindole. Res, resveratrol; Adm, adriamycin; Bai, biacalin. Download figure Download PowerPoint Conclusion Using a 20 nm nanopipette decorated with the specific G4 DNA sequence, this study presented a supersmall electrochemical nanosensor capable of sensitive intracellular K+ detection down to the single-cell level. K+-dependent packing of G4 DNA strands could be sensibly transduced by the small orifice. Exemplified by the primary stage of AVD, changes of intracellular K+ concentrations were monitored after drug treatment. This nanosensor should principally enable the study of stimulus-induced K+ fluctuation in single cells and evaluation of corresponding drugs and therapies. This study is envisioned to inspire more interest in the arenas of intracellular ion detection and single-cell electroanalysis.49–59 Future studies will address equipping such nanodevices with multiple three-dimensional (3D) nanopositioning systems to probe the K+ distribution in neurons.60 Supporting Information Supporting Information is available. Conflict of Interest There is no conflict of interest to report. Funding Information This study was supported by the National Natural Science Foundation of China (grant nos. 21327902, 21675080, and 21974059), the Natural Science Foundation of Jiangsu Province (grant no. BK20170073), and the Excellent Research Program of Nanjing University (no. ZYJH004). Acknowledgments The authors thank the Analytical Instrumentation Center of Nanjing University for analytical tests. References 1. Dubyak G. R. Ion Homeostasis, Channels, and Transporters: An Update on Cellular Mechanisms.Adv. Physiol. Educ.2004, 28, 143−154. Google Scholar 2. Jansson B. Potassium, Sodium, and Cancer: A Review.J. Environ. Pathol. Toxicol. Oncol.1996, 15, 65−73. Google Scholar 3. Eil R.; Vodnala S. K.; Clever D.; Klebanoff C. A.; Sukumar M.; Pan J. H.; Palmer D. C.; Gros A.; Yamamoto T. N.; Patel S. J.; Guittard G. C.; Yu Z.; Carbonaro V.; Okkenhaug K.; Schrump D. S.; Linehan W. M.; Roychoudhuri R.; Restifo N. P. Ionic Immune Suppression within the Tumour Microenvironment Limits T Cell Effector Function.Nature2016, 537, 539−543. Google Scholar 4. Palmer B. F. Regulation of Potassium Homeostasis.Clin. J. Am. Soc. Nephrol.2015, 10, 1050−1060. Google Scholar 5. Sica D. A.; Struthers A. D.; Cushman W. C.; Wood M.; Banas J. S.; Epstein M.Importance of Potassium in Cardiovascular Disease.J. Clin. Hypertens.2002, 4, 198−206. Google Scholar 6. Warny M.; Kelly C. P.Monocytic Cell Necrosis Is Mediated by Potassium Depletion and Caspase−Like Proteases.Am. J. Physiol.1999, 276, C717−C724. Google Scholar 7. Bortner C. D.; Sifre M. I.; Cidlowski J. A.Cationic Gradient Reversal and Cytoskeleton-Independent Volume Regulatory Pathways Define an Early Stage of Apoptosis.J. Biol. Chem.2008, 283, 7219−7229. Google Scholar 8. Reinert M.; Khaldi A.; Zauner A.; Doppenberg E.; Choi S.; Bullock R.High Level of Extracellular Potassium and Its Correlates After Severe Human Head Injury: Relationship to High Intracranial Pressure.J. Neurosurg.2000, 93, 800−807. Google Scholar 9. Bühlmann P.; Pretsch E.; Bakker E.Carrier-Based Ion-Selective Electrodes and Bulk Optodes. 2. Ionophores for Potentiometric and Optical Sensors.Chem. Rev.1998, 98, 1593−1688. Google Scholar 10. Moody G. J.; Thomas J. D. R.Development and Publication of Work with Selective Ion-Sensitive Electrodes.Talanta1972, 19, 623−639. Google Scholar 11. Khalil S. A. H.; Moody G. J.; Thomas J. D. R.Ion-Selective Electrode Determination of Sodium and Potassium in Blood and Urine.Anal. Lett.1986, 19, 1809−1830. Google Scholar 12. Smith P. J. S.Non-Invasive Ion Probes-Tools for Measuring Transmembrane Ion Flux.Nature1995, 378, 645−646. Google Scholar 13. Smith P. J. S.; Trimarchi J.Noninvasive Measurement of Hydrogen and Potassium Ion Flux from Single Cells and Epithelial Structures.Am. J. Physiol. Cell. Physiol.2001, 280, C1−C11. Google Scholar 14. Kang T. M.; Markin V. S.; Hilgemann D. W.Ion Fluxes in Giant Excised Cardiac Membrane Patches Detected and Quantified with Ion-Selective Microelectrodes.J. Gen. Physiol.2003, 121, 325−347. Google Scholar 15. Musa Aziz R.; Boron W. F.; Parker M. D.Using Fluorometry and Ion-Sensitive Microelectrodes to Study the Functional Expression of Heterologously-Expressed Ion Channels and Transporters in Xenopus Oocytes.Methods2010, 51, 134−145. Google Scholar 16. Ali S. M. U.; Asif M. H.; Fulati A.; Nur O.; Willander M.; Brännmark C.; Strålfors P.; Englund U. H.; Elinder F.; Danielsson B.Intracellular K+ Determination with a Potentiometric Microelectrode Based on Zno Nanowires.IEEE Trans. Nanotechnol.2011, 10, 913−919. Google Scholar 17. Takami T.; Iwata F.; Yamazaki K.; Son J. W.; Lee J. K.; Park B. H.; Kawai T.Direct Observation of Potassium Ions in Hela Cell with Ion-Selective Nanopipette Probe.J. Appl. Phys.2012, 111, 044702. Google Scholar 18. Liu Q.; Xiao K.; Wen L.; Lu H.; Liu Y.; Kong X. Y.; Xie G.; Zhang Z.; Bo Z.; Jiang L.Engineered Ionic Gates for Ion Conduction Based on Sodium and Potassium Activated Nanochannels.J. Am. Chem. Soc.2015, 137, 11976−11983. Google Scholar 19. Fang R. C.; Zhang H. C.; Yang L. L.; Wang H. T.; Tian Y.; Zhang X.; Jiang L.Supramolecular Self-Assembly Induced Adjustable Multiple Gating States of Nanofluidic Diodes.J. Am. Chem. Soc.2016, 138, 16372−16379. Google Scholar 20. Acar E. T.; Buchsbaum S. F.; Combs C.; Fornasiero F.; Siwy Z. S.Biomimetic Potassium-Selective Nanopores.Sci. Adv.2019, 5, eaav2568. Google Scholar 21. Xin P. Y.; Kong H. Y.; Sun Y. H.; Zhao L. Y.; Fang H. D.; Zhu H. F.; Jiang T.; Guo J. J.; Zhang Q.; Dong W. P.; Chen C. P.Artificial K+ Channels Formed by Pillararene-Cyclodextrin Hybrid Molecules: Tuning Cation Selectivity and Generating Membrane Potential.Angew. Chem. Int. Ed.2019, 58, 2779−2784. Google Scholar 22. Shen Y.; Wu S. Y.; Rancic V.; Aggarwal A.; Qian Y.; Miyashita S. I.; Ballanyi K.; Campbell R. E.; Dong M.Genetically Encoded Fluorescent Indicators for Imaging Intracellular Potassium Ion Concentration.Commun. Biol.2019, 2, 18. Google Scholar 23. Liu J.; Pan L.; Shang C.; Lu B.; Wu R.; Feng Y.; Chen W.; Zhang R.; Bu J.; Xiong Z.; Bu W.; Du J.; Shi J.A Highly Sensitive and Selective Nanosensor for Near-Infrared Potassium Imaging.Sci. Adv.2020, 6, eaax9757. Google Scholar 24. Pan R. R.; Xu M. C.; Jiang D. C.; Burgess J. D.; Chen H. Y.Nanokit for Single-Cell Electrochemical Analyses.Proc. Natl. Acad. Sci. U. S. A.2016, 113, 11436−11440. Google Scholar 25. Song J.; Xu C. H.; Huang S. Z.; Lei W.; Ruan Y. F.; Lu H. J.; Zhao W.; Xu J. J.; Chen H. Y.Ultrasmall Nanopipette: Toward Continuous Monitoring of Redox Metabolism at Subcellular Level.Angew. Chem. Int. Ed.2018, 57, 13226−13230. Google Scholar 26. Nguyen T. D.; Song M. S.; Ly N. H.; Lee S. Y.; Joo S. W.Nanostars on Nanopipette Tips: A Raman Probe for Quantifying Oxygen Levels in Hypoxic Single Cells and Tumours.Angew. Chem. Int. Ed.2019, 58, 2710−2714. Google Scholar 27. Nadappuram B. P.; Cadinu P.; Barik A.; Ainscough A. J.; Devine M. J.; Kang M.; Gonzalez-Garcia J.; Kittler J. T.; Willison K. R.; Vilar R.; Actis P.; Wojciak Stothard B.; Oh S. H.; Ivanov A. P.; Edel J. B.Nanoscale Tweezers for Single-Cell Biopsies.Nat. Nanotechnol.2019, 14, 80−88. Google Scholar 28. Ying Y. L.; Hu Y. X.; Gao R.; Yu R. J.; Gu Z.; Lee L. P.; Long Y. T.Asymmetric Nanopore Electrode-Based Amplification for Electron Transfer Imaging in Live Cells.J. Am. Chem. Soc.2018, 140, 5385−5392. Google Scholar 29. Liu H.; Jiang Q.; Pang J.; Jiang Z.; Cao J.; Ji L.; Xia X.; Wang K.A Multiparameter pH-Sensitive Nanodevice Based on Plasmonic Nanopores.Adv. Funct. Mater.2018, 28, 1703847. Google Scholar 30. Ozel R. E.; Bulbul G.; Perez J.; Pourmand N.Functionalized Quartz Nanopipette for Intracellular Superoxide Sensing: A Tool for Monitoring Reactive Oxygen Species Levels in Single Living Cell.ACS Sens.2018, 3, 1316−1321. Google Scholar 31. Nascimento R. A. S.; Özel R. E.; Mak W. H.; Mulato M.; Singaram B.; Pourmand N.Single Cell "Glucose Nanosensor" Verifies Elevated Glucose Levels in Individual Cancer Cells.Nano Lett.2016, 16, 1194−1200. Google Scholar 32. Actis P.; Tokar S.; Clausmeyer J.; Babakinejad B.; Mikhaleva S.; Cornut R.; Takahashi Y.; López Córdoba A.; Novak P.; Shevchuck A. I.; Dougan J. A.; Kazarian S. G.; Gorelkin P. V.; Erofeev A. S.; Yaminsky I. V.; Unwin P. R.; Schuhmann W.; Klenerman D.; Rusakov D. A.; Sviderskaya E. V.; Korchev Y. E.Electrochemical Nanoprobes for Single-Cell Analysis.ACS Nano2014, 8, 875−884. Google Scholar 33. Parkinson G. N.; Lee M. P. H.; Neidle S.Crystal Structure of Parallel Quadruplexes from Human Telomeric DNA.Nature2002, 417, 876−880. Google Scholar 34. Huang R. H.; Faber M. K.; Moeggenborg K. J.; Ward D. L.; Dye J. L.Structure of K+(Cryptand[2.2.2J]) Electride and Evidence for Trapped Electron Pairs.Nature1988, 331, 599−601. Google Scholar 35. Xu Y.; Noguchi Y.; Sugiyama H.The New Models of the Human Telomere d[AGGG(TTAGGG)3] in K+ Solution.Bioorg. Med. Chem.2006, 14, 5584−5591. Google Scholar 36. Deore P. S.; Gray M. D.; Chung A. J.; Manderville R. A.Ligand-Induced G-Quadruplex Polymorphism: A DNA Nanodevice for Label-Free Aptasensor Platforms.J. Am. Chem. Soc.2019, 141, 14288−14297. Google Scholar 37. Nicoludis J. M.; Barrett S. P.; Mergny J. L.; Yatsunyk L. A.Interaction of Human Telomeric DNA with n-Methyl Mesoporphyrin IX.Nucleic Acids Res.2012, 40, 5432–5447. Google Scholar 38. Zhao X. P.; Liu F. F.; Hu W. C.; Younis M. R.; Wang C.; Xia X. H.Biomimetic Nanochannel-Ionchannel Hybrid for Ultrasensitive and Label-Free Detection of Microrna in Cells.Anal. Chem.2019, 91, 3582−3589. Google Scholar 39. Zhang Y. J.; Clausmeyer J.; Babakinejad B.; López Córdoba A.; Ali T.; Shevchuk A.; Takahashi Y.; Novak P.; Edwards C.; Lab M.; Gopal S.; Chiappini C.; Anand U.; Magnani L.; Coombes R. C.; Gorelik J.; Matsue T.; Schuhmann W.; Klenerman D.; Sviderskaya E. V.; Korchev Y.Spearhead Nanometric Field-Effect Transistor Sensors for Single-Cell Analysis.ACS Nano2016, 10, 3214−3221. Google Scholar 40. Hamieh M.; Dobrin A.; Cabriolu A.; van der Stegen S. J. C.; Giavridis T.; Mansilla Soto J.; Eyquem J.; Zhao Z.; Whitlock B. M.; Miele M. M.; Li Z.; Cunanan K. M.; Huse M.; Hendrickson R. C.; Wang X.; Rivière I.; Sadelain M.Car T Cell Trogocytosis and Cooperative Killing Regulate Tumour Antigen Escape.Nature2019, 568, 112−116. Google Scholar 41. Tian M. G.; Sun J.; Dong B. L.; Lin W. Y.Dynamically Monitoring Cell Viability in a Dual-Color Mode: Construction of an Aggregation/Monomer-Based Probe Capable of Reversible Mitochondria-Nucleus Migration.Angew. Chem. Int. Ed.2018, 57, 16506−16510. Google Scholar 42. Crowley L. C.; Marfell B. J.; Waterhouse N. J.Analyzing Cell Death by Nuclear Staining with Hoechst 33342.Cold Spring Harb. Protoc.2016, 9, prot087205. Google Scholar 43. Tian Z. Y.; Wang J. H.; Xu M.; Wang Y.; Zhang M.; Zhou Y. Y.Resveratrol Improves Cognitive Impairment by Regulating Apoptosis and Synaptic Plasticity in Streptozotocin-Induced Diabetic Rats.Cell Physiol. Biochem.2016, 40, 1670−1677. Google Scholar 44. Yeh I.; von Deimling A.; Bastian B. C.Clonal BRAF Mutations in Melanocytic Nevi and Initiating Role of BRAF in Melanocytic Neoplasia.J. Natl. Cancer Inst.2013, 105, 917−919. Google Scholar 45. Li L.; Li P.; Fang J.; Li Q.; Xiao H.; Zhou H.; Tang B.Simultaneous Quantitation of Na+ and K+ in Single Normal and Cancer Cells Using a New Near-Infrared Fluorescent Probe.Anal. Chem.2015, 87, 6057−6063. Google Scholar 46. Bilim V. N.; Tomita Y.; Kawasaki T.; Takeda M.; Takahashi K.Adriamycin (ADM) Induced Apoptosis in Transitional Cell Cancer (TCC) Cell Lines Accompanied by p21 WAF1/CIP1 Induction.Apoptosis1997, 2, 207−213. Google Scholar 47. Wang L. H.; Yang J. Y.; Yang S. N.; Li Y.; Ping G. F.; Hou Y.; Cui W.; Wang Z. Z.; Xiao W.; Wu C. F.Suppression of NF-kB Signaling and P-Glycoprotein Function by Gambogic Acid Synergistically Potentiates Adriamycin-Induced Apoptosis in Lung Cancer.Curr. Cancer Drug Targets2014, 14, 91−103. Google Scholar 48. Shu Y. J.; Bao R. F.; Wu X. S.; Weng H.; Ding Q.; Cao Y.; Li M. L.; Mu J. S.; Wu W. G.; Ding Q. C.; Liu T. Y.; Jiang L.; Hu Y. P.; Tan Z. J.; Wang P.; Liu Y. B.Baicalin Induces Apoptosis of Gallbladder Carcinoma Cells in Vitro via a Mitochondrial-Mediated Pathway and Suppresses Tumor Growth in Vivo.Anticancer Agents Med. Chem.2014, 14, 1136−1145. Google Scholar 49. Son D.; Park S. Y.; Kim B.; Koh J. T.; Kim T. H.; An S.; Jang D.; Kim G. T.; Jhe W.; Hong S.Nanoneedle Transistor-Based Sensors for the Selective Detection of Intracellular Calcium Ions.ACS Nano2011, 5, 3888−3895. Google Scholar 50. Li X.; Majdi S.; Dunevall J.; Fathali H.; Ewing A. G.Quantitative Measurement of Transmitters in Individual Vesicles in the Cytoplasm of Single Cells with Nanotip Electrodes.Angew. Chem. Int. Ed.2015, 54, 11978–11982. Google Scholar 51. Li X.; Dunevall J.; Ewing A. G.Quantitative Chemical Measurements of Vesicular Transmitters with Electrochemical Cytometry.Acc. Chem. Res.2016, 49, 2347–2354. Google Scholar 52. Zhang X.W.; Qiu Q. F.; Jiang H.; Zhang F. L.; Liu Y. L.; Amatore C.; Huang W. H.Real-Time Intracellular Measurements of ROS and RNS in Living Cells with Single Core-Shell Nanowire Electrodes.Angew. Chem. Int. Ed.2017, 56, 12997–13000. Google Scholar 53. Li Y.; Hu K.; Yu Y.; Rotenberg S. A.; Amatore C.; Mirkin M. V.Direct Electrochemical Measurements of Reactive Oxygen and Nitrogen Species in Nontransformed and Metastatic Human Breast Cells.J. Am. Chem. Soc.2017, 139, 13055−13062. Google Scholar 54. Pan R. R.; Xu M. C.; Burgess J. D.; Jiang D. C.; Chen H. Y.Direct Electrochemical Observation of Glucosidase Activity in Isolated Single Lysosomes from a Living Cell.Proc. Natl. Acad. Sci. U. S. A.2018, 115, 4087−4092. Google Scholar 55. Hu K.; Li Y.; Rotenberg S. A.; Amatore C.; Mirkin M. V.Electrochemical Measurements of Reactive Oxygen and Nitrogen Species Inside Single Phagolysosomes of Living Macrophages.J. Am. Chem. Soc.2019, 141, 4564–4568. Google Scholar 56. Yu R. J.; Ying Y. L.; Gao R.; Long Y. T.Confined Nanopipette Sensing: From Single Molecules, Single Nanoparticles, to Single Cells.Angew. Chem. Int. Ed.2019, 58, 3706−3714. Google Scholar 57. Zhu W.; Gu C.; Dunevall J.; Ren L.; Zhou X.; Ewing A. G.Combined Amperometry and Electrochemical Cytometry Reveal Differential Effects of Cocaine and Methylphenidate on Exocytosis and the Fraction of Chemical Release.Angew. Chem. Int. Ed.2019, 58, 4238–4242. Google Scholar 58. Zhang X. W.; Oleinick A.; Jiang H.; Liao Q. L.; Qiu Q. F.; Svir I.; Liu Y. L.; Amatore C.; Huang W. H.Electrochemical Monitoring of ROS/RNS Homeostasis Within Individual Phagolysosomes Inside Single Macrophages.Angew. Chem. Int. Ed.2019, 58, 7753–7756. Google Scholar 59. Wang D.; Qi G.; Zhou Y.; Zhang Y.; Wang B.; Hu P.; Jin Y.Single-Cell ATP Detection and Content Analyses in Electrostimulus-Induced Apoptosis Using Functionalized Glass Nanopipettes.Chem. Commun.2020, 56, 1561−1564. Google Scholar 60. Padmawar P.; Yao X. M.; Bloch O.; Manley G. T.; Verkman A. S.K+ Waves in Brain Cortex Visualized Using a Long-Wavelength K+-Sensing Fluorescent Indicator.Nat. Methods2005, 2, 825−827. Google Scholar Previous articleNext article FiguresReferencesRelatedDetails Issue AssignmentVolume 3Issue 9Page: 2359-2367Supporting Information Copyright & Permissions© 2020 Chinese Chemical SocietyKeywordsnanosensortransmembranepotassium ionssingle-cell analysisG-quadruplex DNAAcknowledgmentsThe authors thank the Analytical Instrumentation Center of Nanjing University for analytical tests. Downloaded 1,220 times PDF DownloadLoading ...
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