Microimaging of Oxygen Concentration Near Live Photosynthetic Cells by Electron Spin Resonance

Overview

Journal Biophys J

Publisher Cell Press

Specialty Biophysics

Date 2010 Aug 5

PMID 20682276

Citations 9

Authors

Revital Halevy

Victor Tormyshev

Aharon Blank

Affiliations

Soon will be listed here.

Abstract

We present what is, to our knowledge, a new methodology for high-resolution three-dimensional imaging of oxygen concentration near live cells. The cells are placed in the buffer solution of a stable paramagnetic probe, and electron spin-resonance microimaging is employed to map out the probe's spin-spin relaxation time (T(2)). This information is directly linked to the concentration of the oxygen molecule. The method is demonstrated with a test sample and with a small amount of live photosynthetic cells (cyanobacteria), under conditions of darkness and light. Spatial resolution of approximately 30 x 30 x 100 microm is demonstrated, with approximately microM oxygen concentration sensitivity and sub-fmol absolute oxygen sensitivity per voxel. The use of electron spin-resonance microimaging for oxygen mapping near cells complements the currently available techniques based on microelectrodes or fluorescence/phosphorescence. Furthermore, with the proper paramagnetic probe, it will also be readily applicable for intracellular oxygen microimaging, a capability which other methods find very difficult to achieve.

Citing Articles

Rapid Scan EPR Oxygen Imaging in Photoactivated Resin Used for Stereolithographic 3D Printing.

Tseytlin O, OConnell R, Sivashankar V, Bobko A, Tseytlin M 3D Print Addit Manuf. 2022; 8(6):358-365.

PMID: 34977276 PMC: 8713732. DOI: 10.1089/3dp.2020.0170.

Triarylmethyl Radicals: EPR Study of C Hyperfine Coupling Constants.

Kuzhelev A, Tormyshev V, Rogozhnikova O, Trukhin D, Troitskaya T, Strizhakov R Z Phys Chem (N F). 2017; 231(4):777-794.

PMID: 28539703 PMC: 5439964. DOI: 10.1515/zpch-2016-0811.

Rapid Scan Electron Paramagnetic Resonance Opens New Avenues for Imaging Physiologically Important Parameters In Vivo.

Biller J, Mitchell D, Tseytlin M, Elajaili H, Rinard G, Quine R J Vis Exp. 2016; (115).

PMID: 27768025 PMC: 5092072. DOI: 10.3791/54068.

Facile and High-Yielding Synthesis of TAM Biradicals and Monofunctional TAM Radicals.

Trukhin D, Rogozhnikova O, Troitskaya T, Vasiliev V, Bowman M, Tormyshev V Synlett. 2016; 27(6):893-899.

PMID: 27065567 PMC: 4826066. DOI: 10.1055/s-0035-1561299.

Preparation of Diversely Substituted Triarylmethyl Radicals by the Quenching of Tris(2,3,5,6-tetrathiaaryl)methyl Cations with C-, N-, P-, and S-Nucleophiles.

Tormyshev V, Rogozhnikova O, Bowman M, Trukhin D, Troitskaya T, Vasiliev V European J Org Chem. 2014; 2014(2):371-380.

PMID: 24883040 PMC: 4038673. DOI: 10.1002/ejoc.201301161.

References

Mancini D, Wilson J, Bolinger L, Li H, Kendrick K, Chance B . In vivo magnetic resonance spectroscopy measurement of deoxymyoglobin during exercise in patients with heart failure. Demonstration of abnormal muscle metabolism despite adequate oxygenation. Circulation. 1994; 90(1):500-8. DOI: 10.1161/01.cir.90.1.500. View

Land S, Porterfield D, Sanger R, Smith P . The self-referencing oxygen-selective microelectrode: detection of transmembrane oxygen flux from single cells. J Exp Biol. 1998; 202(Pt 2):211-8. DOI: 10.1242/jeb.202.2.211. View

Finikova O, Lebedev A, Aprelev A, Troxler T, Gao F, Garnacho C . Oxygen microscopy by two-photon-excited phosphorescence. Chemphyschem. 2008; 9(12):1673-9. PMC: 2645351. DOI: 10.1002/cphc.200800296. View

Zhang G, Palmer G, Dewhirst M, Fraser C . A dual-emissive-materials design concept enables tumour hypoxia imaging. Nat Mater. 2009; 8(9):747-51. PMC: 2846459. DOI: 10.1038/nmat2509. View

Sud D, Mehta G, Mehta K, Linderman J, Takayama S, Mycek M . Optical imaging in microfluidic bioreactors enables oxygen monitoring for continuous cell culture. J Biomed Opt. 2006; 11(5):050504. DOI: 10.1117/1.2355665. View

Tang X, Chisholm D, Dismukes G, Brudvig G, Diner B . Spectroscopic evidence from site-directed mutants of Synechocystis PCC6803 in favor of a close interaction between histidine 189 and redox-active tyrosine 160, both of polypeptide D2 of the photosystem II reaction center. Biochemistry. 1993; 32(49):13742-8. DOI: 10.1021/bi00212a045. View

Xi Q, Cheranov S, Jaggar J . Mitochondria-derived reactive oxygen species dilate cerebral arteries by activating Ca2+ sparks. Circ Res. 2005; 97(4):354-62. PMC: 1352308. DOI: 10.1161/01.RES.0000177669.29525.78. View

Owenius R, Eaton G, Eaton S . Frequency (250 MHz to 9.2 GHz) and viscosity dependence of electron spin relaxation of triarylmethyl radicals at room temperature. J Magn Reson. 2004; 172(1):168-75. DOI: 10.1016/j.jmr.2004.10.007. View

Yong L, Harbridge J, Quine R, Rinard G, Eaton S, Eaton G . Electron spin relaxation of triarylmethyl radicals in fluid solution. J Magn Reson. 2001; 152(1):156-61. DOI: 10.1006/jmre.2001.2379. View

10.

Swartz H . Measuring real levels of oxygen in vivo: opportunities and challenges. Biochem Soc Trans. 2002; 30(2):248-52. DOI: 10.1042/. View

11.

Springett R, Swartz H . Measurements of oxygen in vivo: overview and perspectives on methods to measure oxygen within cells and tissues. Antioxid Redox Signal. 2007; 9(8):1295-301. DOI: 10.1089/ars.2007.1620. View

12.

Schledz M, Seidler A, Beyer P, Neuhaus G . A novel phytyltransferase from Synechocystis sp. PCC 6803 involved in tocopherol biosynthesis. FEBS Lett. 2001; 499(1-2):15-20. DOI: 10.1016/s0014-5793(01)02508-x. View

13.

Ardenkjaer-Larsen J, Laursen I, Leunbach I, Ehnholm G, Wistrand L, Petersson J . EPR and DNP properties of certain novel single electron contrast agents intended for oximetric imaging. J Magn Reson. 1998; 133(1):1-12. DOI: 10.1006/jmre.1998.1438. View

14.

Blank A, Dunnam C, Borbat P, Freed J . High resolution electron spin resonance microscopy. J Magn Reson. 2003; 165(1):116-27. DOI: 10.1016/s1090-7807(03)00254-4. View

15.

Zweier J, Kuppusamy P . In vivo EPR spectroscopy of free radicals in the heart. Environ Health Perspect. 1994; 102 Suppl 10:45-51. PMC: 1566969. DOI: 10.1289/ehp.94102s1045. View

16.

Swartz H . Using EPR to measure a critical but often unmeasured component of oxidative damage: oxygen. Antioxid Redox Signal. 2004; 6(3):677-86. DOI: 10.1089/152308604773934440. View

17.

Kuhl M, Holst G, Larkum A, Ralph P . IMAGING OF OXYGEN DYNAMICS WITHIN THE ENDOLITHIC ALGAL COMMUNITY OF THE MASSIVE CORAL PORITES LOBATA(1). J Phycol. 2016; 44(3):541-50. DOI: 10.1111/j.1529-8817.2008.00506.x. View

18.

Blank A, Suhovoy E, Halevy R, Shtirberg L, Harneit W . ESR imaging in solid phase down to sub-micron resolution: methodology and applications. Phys Chem Chem Phys. 2009; 11(31):6689-99. DOI: 10.1039/b905943a. View

19.

Kutala V, Parinandi N, Pandian R, Kuppusamy P . Simultaneous measurement of oxygenation in intracellular and extracellular compartments of lung microvascular endothelial cells. Antioxid Redox Signal. 2004; 6(3):597-603. DOI: 10.1089/152308604773934350. View

20.

Liu K, Gast P, Moussavi M, Norby S, Vahidi N, Walczak T . Lithium phthalocyanine: a probe for electron paramagnetic resonance oximetry in viable biological systems. Proc Natl Acad Sci U S A. 1993; 90(12):5438-42. PMC: 46735. DOI: 10.1073/pnas.90.12.5438. View