Nanoscale detection and real-time monitoring of free radicals in a single living cell under the stimulation of targeting moieties using a nanodiamond quantum sensor

Kaiqi Wu; Qi Lu; Maabur Sow; Priyadharshini Balasubramanian; Fedor Jelezko; Tanja Weil; Yingke Wu

doi:10.1080/26941112.2024.2336524

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Nanoscale detection and real-time monitoring of free radicals in a single living cell under the stimulation of targeting moieties using a nanodiamond quantum sensor

Research Article | Updated：2025-01-22

- Nanoscale detection and real-time monitoring of free radicals in a single living cell under the stimulation of targeting moieties using a nanodiamond quantum sensor
- Functional Diamond Vol. 4, Issue 1, (2024)
- Affiliations：
  
  1. a Max Planck Institute for Polymer Research
  2. b Institute for Quantum Optics and Center for Integrated Quantum Science and Technology (IQST) Ulm University
- Author bio：
- Funds：
- DOI：10.1080/26941112.2024.2336524
  CLC：
- Online First：08 April 2024，
  
  Published：2024
- Accepted：
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Kaiqi Wu, Qi Lu, Maabur Sow, et al. Nanoscale detection and real-time monitoring of free radicals in a single living cell under the stimulation of targeting moieties using a nanodiamond quantum sensor[J]. Functional Diamond2024, 4(1).
DOI：

Kaiqi Wu, Qi Lu, Maabur Sow, et al. Nanoscale detection and real-time monitoring of free radicals in a single living cell under the stimulation of targeting moieties using a nanodiamond quantum sensor[J]. Functional Diamond2024, 4(1). DOI： 10.1080/26941112.2024.2336524.

摘要

Abstract

Intracellular radicals play important roles in cell signaling and regulation of growth factors

cytokines

transcription

apoptosis

and immunomodulation

among others. To gain a more comprehensive understanding of their biological functions from a spatio-temporal perspective

there is a great need for nanoscale sensitive tools that allow real-time detection of these reactive species. Currently

intracellular radical probes are based on chemical reactions that could significantly alter radical levels during detection. Due to the excellent biocompatibility and favorable photophysical properties of nitrogen-vacancy (NV

–

) centers in fluorescent nanodiamonds (fNDs)

the fNDs can serve as a powerful and chemically inert nanotool for intracellular radical detection. In this study

a positively charged nanogel (NG) coating was prepared to prevent the precipitation of fNDs and promote cellular internalization. After internalization of nanodiamond-nanogels (fND-NGs)

different stimulators

namely somatostatin (SST)

triphenylphosphonium (TPP)

and trans-activator of transcription

(TAT) peptide

which are widely used cell- or organelle-targeting ligands in medicine

drug delivery

and diagnostics

were applied to stimulate the cells. In parallel

the intracellular radical changes under stimulation of SST

TPP

and TAT ligands were monitored by fND-NGs in a home-built optically detected magnetic resonance (ODMR) microscope. Our method allows for detecting intracellular radicals

in-situ

and monitoring their real-time changes during incubation with the targeting ligands in a single living cell. We believe that our method will provide insights into the generation of radical stress in cells

which could improve our fundamental understanding of the pharmacology and signaling pathways of widely used cell- and organelle-targeting ligands associated with free radicals.

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Keywords

references

Pizzino G, Irrera N, Cucinotta M, et al. Oxidative stress: harms and benefits for human health. Oxidative Med Cell Longev. 2017;2017:1–13. (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Lobo V, Patil A, Phatak A, et al. Free radicals, antioxidants and functional foods: impact on human health. Phcog Rev. 2010;4(8):118–126. (Open in a new window)Google Scholar

Okada F. Inflammation and free radicals in tumor development and progression. Redox Rep. 2002;7(6):357–368. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Nathan C, Cunningham-Bussel A. Beyond oxidative stress: an immunologist’s guide to reactive oxygen species. Nat Rev Immunol. 2013;13(5):349–361. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Radi R. Oxygen radicals, nitric oxide, and peroxynitrite: redox pathways in molecular medicine. Proc Natl Acad Sci USA. 2018;115(23):5839–5848. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Knight JA. Review: free radicals, antioxidants, and the immune system. Ann Clin Lab Sci. 2000;30(2):145–158. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Andrés C, Pérez de la Lastra J, Juan C, et al. The role of reactive species on innate immunity. Vaccines. 2022;10(10):1735. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Gomes A, Fernandes E, Lima JL. Fluorescence probes used for detection of reactive oxygen species. J Biochem Biophys Methods. 2005;65(2–3):45–80. (Open in a new window)PubMed(Open in a new window)Google Scholar

Damle VG, Wu K, Arouri DJ, et al. Detecting free radicals post viral infections. Free Radic Biol Med. 2022;191:8–23. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Kalyanaraman B, Darley-Usmar V, Davies KJ, et al. Measuring reactive oxygen and nitrogen species with fluorescent probes: challenges and limitations. Free Radic Biol Med. 2012;52(1):1–6. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Wu Y, Weil T. Recent developments of nanodiamond quantum sensors for biological applications. Adv Sci. 2022;9(19):2200059. (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Schirhagl R, Chang K, Loretz M, et al. Nitrogen-vacancy centers in diamond: nanoscale sensors for physics and biology. Annu Rev Phys Chem. 2014;65(1):83–105. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Steinert S, Ziem F, Hall LT, et al. Magnetic spin imaging under ambient conditions with sub-cellular resolution. Nat Commun. 2013;4(1):1607. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Vaijayanthimala V, Tzeng Y, Chang H, et al. The biocompatibility of fluorescent nanodiamonds and their mechanism of cellular uptake. Nanotechnology. 2009;20(42):425103. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Barton J, Gulka M, Tarabek J, et al. Nanoscale dynamic readout of a chemical redox process using radicals coupled with nitrogen-vacancy centers in nanodiamonds. ACS Nano. 2020;14(10):12938–12950. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Wu Y, Balasubramanian P, Wang Z, et al. Detection of few hydrogen peroxide molecules using self-reporting fluorescent nanodiamond quantum sensors. J Am Chem Soc. 2022;144(28):12642–12651. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Maze JR, Stanwix PL, Hodges JS, et al. Nanoscale magnetic sensing with an individual electronic spin in diamond. Nature. 2008;455(7213):644–647. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Ackermann K, Wort JL, Bode BE. Pulse dipolar EPR for determining nanomolar binding affinities. Chem Commun. 2022;58(63):8790–8793. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Sigaeva A, Shirzad H, Martinez FP, et al. Diamond-based nanoscale quantum relaxometry for sensing free radical production in cells. Small. 2022;18(44):2105750. (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Sigaeva A, Norouzi N, Schirhagl R. Intracellular relaxometry, challenges, and future directions. ACS Cent Sci. 2022;8(11):1484–1489. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Nie L, Nusantara AC, Damle VG, et al. Quantum monitoring of cellular metabolic activities in single mitochondria. Sci Adv. 2021;7(21):eabf0573. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Wu K, Vedelaar TA, Damle VG, et al. Applying NV center-based quantum sensing to study intracellular free radical response upon viral infections. Redox Biol. 2022;52:102279. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Wu K, Nie L, Nusantara AC, et al. Diamond relaxometry as a tool to investigate the free radical dialogue between macrophages and bacteria. ACS Nano. 2023;17(2):1100–1111. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Lu Q, Vosberg B, Wang Z, et al. Unraveling eumelanin radical formation by nanodiamond optical relaxometry in a living cell. J. Am. Chem. Soc. 2024;146(11):7222–7232. (Open in a new window)Google Scholar

Wu Y, Alam MNA, Balasubramanian P, et al. Nanodiamond theranostic for light-controlled intracellular heating and nanoscale temperature sensing. Nano Lett. 2021;21(9):3780–3788. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Wu Y, Alam MNA, Balasubramanian P, et al. Fluorescent nanodiamond–nanogels for nanoscale sensing and photodynamic applications. Adv Biomed Res. 2021;1(7):2000101. (Open in a new window)Google Scholar

Theodoropoulou M, Stalla GK. Somatostatin receptors: from signaling to clinical practice. Front Neuroendocrinol. 2013;34(3):228–252. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Zielonka J, Joseph J, Sikora A, et al. Mitochondria-targeted triphenylphosphonium-based compounds: syntheses, mechanisms of action, and therapeutic and diagnostic applications. Chem Rev. 2017;117(15):10043–10120. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Gump JM, Dowdy SF. TAT transduction: the molecular mechanism and therapeutic prospects. Trends Mol Med. 2007;13(10):443–448. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Jung HS, Cho KJ, Seol Y, et al. Polydopamine encapsulation of fluorescent nanodiamonds for biomedical applications. Adv Funct Mater. 2018;28(33):1801252. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Shamsi BH, Chatoo M, Xu X, et al. Versatile functions of somatostatin and somatostatin receptors in the gastrointestinal system. Front Endocrinol. 2021;12:652363. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Bousquet C, Puente E, Buscail L, et al. Antiproliferative effect of somatostatin and analogs. Chemotherapy. 2001;47(Suppl. 2):30–39. (Open in a new window)PubMed(Open in a new window)Google Scholar

Zou Y, Xiao X, Li Y, et al. Somatostatin analogues inhibit cancer cell proliferation in an SSTR2-dependent manner via both cytostatic and cytotoxic pathways. Oncol Rep. 2009;21(2):379–386. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Sun L, Coy DH. Somatostatin receptor-targeted anti-cancer therapy. CDD. 2011;8(1):2–10. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Grötzinger C, Wiedenmann B. Somatostatin receptor targeting for tumor imaging and therapy. Ann N Y Acad Sci. 2004;1014(1):258–264. (Open in a new window)PubMed(Open in a new window)Google Scholar

Arena S, Pattarozzi A, Corsaro A, et al. Somatostatin receptor subtype-dependent regulation of nitric oxide release: involvement of different intracellular pathways. Mol Endocrinol. 2005;19(1):255–267. (Open in a new window)PubMed(Open in a new window)Google Scholar

Akaike T, Maeda H. Nitric oxide and virus infection. Immunology. 2000;101(3):300–308. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Hou C, Metcalfe NB, Salin K. Is mitochondrial reactive oxygen species production proportional to oxygen consumption? A theoretical consideration. Bioessays. 2021;43(4):e2000165. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Reily C, Mitchell T, Chacko BK, et al. Mitochondrially targeted compounds and their impact on cellular bioenergetics. Redox Biol. 2013;1(1):86–93. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Frankel AD, Pabo CO. Cellular uptake of the TAT protein from human immunodeficiency virus. Cell. 1988;55(6):1189–1193. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Green M, Loewenstein PM. Autonomous functional domains of chemically synthesized human immunodeficiency virus TAT trans-activator protein. Cell. 1988;55(6):1179–1188. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Kuroda Y, Kato-Kogoe N, Tasaki E, et al. Oligopeptides derived from autophosphorylation sites of EGF receptor suppress EGF-stimulated responses in human lung carcinoma a549 cells. Eur J Pharmacol. 2013;698(1–3):87–94. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Ruseska I, Zimmer A. Internalization mechanisms of cell-penetrating peptides. Beilstein J Nanotechnol. 2020;11:101–123. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Sarder P, Nehorai A. Deconvolution methods for 3-D fluorescence microscopy images. IEEE Signal Process Mag. 2006;23(3):32–45. (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Dunn KW, Kamocka MM, McDonald JH. A practical guide to evaluating colocalization in biological microscopy. Am J Physiol-Cell Physiol. 2011;300(4):C723–C742. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Zhang Y, Sharmin R, Sigaeva A, et al. Not all cells are created equal – endosomal escape in fluorescent nanodiamonds in different cells. Nanoscale. 2021;13(31):13294–13300. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Stauffer W, Sheng H, Lim H. Ezcolocalization: an imagej plugin for visualizing and measuring colocalization in cells and organisms. Sci Rep. 2018;8(1):15764. (Open in a new window)PubMed(Open in a new window)Google Scholar

Martens TF, Remaut K, Demeester J, et al. Intracellular delivery of nanomaterials: how to catch endosomal escape in the act. Nano Today. 2014;9(3):344–364. (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Binder JM, Stark A, Tomek N, et al. Qudi: a modular python suite for experiment control and data processing. SoftwareX. 2017;6:85–90. (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Yanagi T, Kaminaga K, Suzuki M, et al. All-optical wide-field selective imaging of fluorescent nanodiamonds in cells, in vivo and ex vivo. ACS Nano. 2021;15(8):12869–12879. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Bluvstein D, Zhang Z, Jayich ACB. Identifying and mitigating charge instabilities in shallow diamond nitrogen-vacancy centers. Phys Rev Lett. 2019;122(7):076101. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Barbosa I, Gutsche J, Widera A. Impact of charge conversion on NV-center relaxometry. Phys Rev B. 2023;108(7):075411. (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Barnett P. Somatostatin and somatostatin receptor physiology. ENDO. 2003;20(3):255–264. (Open in a new window)Google Scholar

Hofland LJ, Lamberts SW. The pathophysiological consequences of somatostatin receptor internalization and resistance. Endocr Rev. 2003;24(1):28–47. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Cakir M, Dworakowska D, Grossman A. Somatostatin receptor biology in neuroendocrine and pituitary tumours: part 1-molecular pathways. J Cellular Molecular Medi. 2010;14(11):2570–2584. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

White RE, Schonbrunn A, Armstrong DL. Somatostatin stimulates Ca2+-activated K+ channels through protein dephosphorylation. Nature. 1991;351(6327):570–573. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Choi EJ, Jeon CH, Lee IK. Ferric ammonium citrate upregulates PD-L1 expression through generation of reactive oxygen species. J Immunol Res. 2022;2022:1–8. (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Zorko M, Langel U. Cell-penetrating peptides: mechanism and kinetics of cargo delivery. Adv Drug Deliv Rev. 2005;57(4):529–545. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

de Guillebon T, Vindolet B, Roch JF, et al. Temperature dependence of the longitudinal spin relaxation time t1 of single nitrogen-vacancy centers in nanodiamonds. Phys Rev B. 2020;102(16):165427. (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Fujisaku T, Tanabe R, Onoda S, et al. pH nanosensor using electronic spins in diamond. ACS Nano. 2019;13(10):11726–11732. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Shi FZ, Zhang Q, Wang PF, et al. Single-protein spin resonance spectroscopy under ambient conditions. Science. 2015;347(6226):1135–1138. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Shi FZ, Kong F, Zhao PJ, et al. Single-DNA electron spin resonance spectroscopy in aqueous solutions. Nat Methods. 2018;15(9):697–699. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

Wood JDA, Tetienne JP, Broadway DA, et al. Microwave-free nuclear magnetic resonance at molecular scales. Nat Commun. 2017;8(1):15950. (Open in a new window)PubMed(Open in a new window)Google Scholar

Qin Z, Wang Z, Kong F, et al. In situ electron paramagnetic resonance spectroscopy using single nanodiamond sensors. Nat Commun. 2023;14(1):6278. (Open in a new window)PubMed (Open in a new window)Web of Science ®(Open in a new window)Google Scholar

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