Singlet Oxygen Microscope:  From Phase-Separated Polymers to Single Biological Cells

Snyder, John W.; Zebger, Ingo; Gao, Zhan; Poulsen, Lars; Frederiksen, Peter; Skovsen, Esben; McIlroy, Sean; Klinger, Markus; Andersen, Lars Klembt; Ogilby, Peter R.

doi:10.1021/ar040075y

Cited by 75 publications

(72 citation statements)

References 42 publications

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“…[12][13][14][15][16] A key aspect of our work is that, using a focused laser beam, sensitizer excitation can be confined to small sub-cellular spatial domains. 15,17,18 Using this approach, it is now possible to provide unique insight into singlet-oxygen-mediated processes that occur in a cell.…”

Section: Introductionmentioning

confidence: 99%

Singlet Oxygen in a Cell: Spatially Dependent Lifetimes and Quenching Rate Constants

2008

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Singlet molecular oxygen, O 2 (a 1 ∆ g ), can be created in a single cell from ground state oxygen, O 2 (X 3 Σ g -), upon focused laser irradiation of an intracellular sensitizer. This cytotoxic species can subsequently be detected by its 1270 nm phosphorescence (a 1 ∆ g → X 3 Σ g -) with subcellular spatial resolution. The singlet oxygen lifetime determines its diffusion distance and, hence, the intracellular volume element in which singlet-oxygen-initiated perturbation of the cell occurs. In this study, the time-resolved phosphorescence of singlet oxygen produced by the sensitizers chlorin (Chl) and 5,10,15,20-tetrakis(N-methyl-4-pyridyl)-21H,23H-porphine (TMPyP) was monitored. These molecules localize in different domains of a living cell. The data indicate that (i) the singlet oxygen lifetime and (ii) the rate constant for singlet oxygen quenching by added NaN 3 depend on whether Chl or TMPyP was the photosensitizer.These observations likely reflect differences in the chemical and physical constituency of a given subcellular domain (e.g., spatially-dependent oxygen and NaN 3 diffusion coefficients) and, as such, are evidence that singlet oxygen responds to the inherent heterogeneity of a cell.Thus, despite a relatively long intracellular lifetime, singlet oxygen does not diffuse a great distance from its site of production. This is a consequence of an apparent intracellular viscosity that is comparatively large.3

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Section: Introductionmentioning

confidence: 99%

Singlet Oxygen in a Cell: Spatially Dependent Lifetimes and Quenching Rate Constants

2008

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“…In particular, the 1 O 2 diffusion in living cells has been a subject of debate for lasted twenty years. Recently, direct microscopic detection of 1 O 2 in a single cell by monitoring its 1270 nm luminescence has been reported [9][10][11][12][13]. Unfortunately, all of these studies require the use of deuterated solvents (i.e.…”

Section: Introductionmentioning

confidence: 99%

Indirect imaging of singlet oxygen generation from a single cell

et al. 2011

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Singlet oxygen (1 O 2 ) can be generated in a living cell upon focused laser irradiation of the intracellular photosensitizers.1 O2 lifetime in the living cells is shortened by the reactions with cellular molecules, and thus the 1 O2 diffusion in a single cell has attracted much attention. In this study, 1 O2 generation from the plasma membrane-targeted protoporphyrin IX (PpIX) and nuclear-targeted meso-Tetra (N-methyl-4-pyridyl) porphine tetra tosylate (TMPyP) in human nasopharyngeal carcinoma CNE2 cells was indirectly imaged by using a fluorescence probe Singlet Oxygen Sensor Green agent (SOSG), respectively. The confocal images indicate that the green fluorescence of SOSG in the vicinity of the PpIX-sensitized cells was dramatically enhanced with the increase of the irradiation time and intracellular PpIX, while there is no significant enhancement for the unsensitized and TMPyP-sensitized cells. The obtained results suggest that the 1 O2 generated from the plasma membrane-targeted PpIX in the CNE2 cells can escape into the extracellular medium and react with the SOSG to produce SOSG endoperoxides (SOSG-EP). Moreover, the fluorescence enhancement of SOSG mainly depends on the subcellular localization and intracellular uptake of the photosensitizers. Depending on the site of 1 O2 generation, 1 O2 generated in the plasma membrane can escape from the cell interior into the extracellular environment, while the 1 O2 generated in the nucleus cannot. Our findings indicate that SOSG holds great promise for the indirect imaging of the 1 O 2 that can escape from single intact living cells. The average intensity of four regions (marked as A, B, C, and D) from special time point was followed in each sample using the image-guided spectral analysis function

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“…Figure 8 shows that this emission is indeed comparable to the emission of certain NIR luminescent lanthanide complexes [18]. The efforts made in the instrumental development of microscopes that image the luminescence of singlet oxygen [88] are therefore of great interest for the imaging of NIR luminescent lanthanide complexes.…”

Section: Detection Of Near-infrared Luminescencementioning

confidence: 73%

“…The challenge of detecting weak NIR (700-1,600 nm) luminescence in organic and biological media is not limited to luminescent lanthanide complexes. In this respect, it is interesting to mention the efforts currently made for the microscopic detection of singlet oxygen phosphorescence [87,88]. Singlet oxygen is a highly reactive photochemical species, usually obtained through the energy transfer from excited triplet states of organic chromophores such as porphyrins.…”

Section: Detection Of Near-infrared Luminescencementioning

confidence: 99%

Near-Infrared Luminescent Labels and Probes Based on Lanthanide Ions and Their Potential for Applications in Bioanalytical Detection and Imaging

Werts

2010

Lanthanide Luminescence

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The use of near-infrared (NIR) light in bioanalytical detection and biological imaging presents certain advantages over UV-visible light in terms of penetration depth, reduction of background signals and decreased phototoxicity. Under suitable conditions, complexes of ytterbium(III)-, neodymium(III)-and erbium (III)-containing organic chromophores may display relatively bright NIR luminescence upon excitation with visible light. This contrasts with the more commonly studied visibly luminescent europium(III) and terbium(III) complexes, which generally need UV or blue light for excitation. We discuss the current knowledge on NIR luminescence of lanthanide complexes (including holmium(III), praseodymium(III) and thulium(III)) in solution. It is suggested that among the NIR luminescent lanthanide complexes, ytterbium(III) complexes are likely to be the most promising candidates for biophotonic applications. The study of NIR luminescence and its application in bioanalytical detection are enabled by progress in detector and light source technology, which are also addressed. Recent developments in lanthanide-based NIR luminescent materials, including complexes and doped nanoparticles, are discussed in the light of their potential for bioanalytical application.

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Singlet Oxygen Microscope: From Phase-Separated Polymers to Single Biological Cells

Cited by 75 publications

References 42 publications

Singlet Oxygen in a Cell: Spatially Dependent Lifetimes and Quenching Rate Constants

Singlet Oxygen in a Cell: Spatially Dependent Lifetimes and Quenching Rate Constants

Indirect imaging of singlet oxygen generation from a single cell

Near-Infrared Luminescent Labels and Probes Based on Lanthanide Ions and Their Potential for Applications in Bioanalytical Detection and Imaging

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