Confocal calcium imaging with ultraviolet laser- scanning microscopy and indo-1

Minamikawa, Tetsuhiro; Takamatsu, Tetsuro; Kashima, Shingo; Fushiki, Shinji; Fujita, Setsuya

doi:10.1016/0968-4328(93)90031-u

Cited by 17 publications

(7 citation statements)

References 16 publications

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“…Although this is not so severe a problem in wide-field microscopy, corrections for chromatic aberrations are crucial for confocal imaging. Consequently, UV-based laser systems require specialized optical components (253,280), which increases its cost and decreases overall signal intensity.…”

Section: B Ultraviolet-wavelength Excitation Fluorescent Indicatorsmentioning

confidence: 99%

“…Because the light sources commonly employed in CLSM are argon or argon-krypton lasers, the usable range of excitation wavelengths is quite restricted. However, recently some UV-excitable CLSM have been produced (253,280), and temporal resolution has been improved to video-frame rate (30 frames/s) or faster (15,136,188,278,392).…”

Section: Confocal Laser Scanning Microscopymentioning

confidence: 99%

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Measurement of Intracellular Calcium

Takahashi

Camacho

Lechleiter

et al. 1999

Physiological Reviews

674

511

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To a certain extent, all cellular, physiological, and pathological phenomena that occur in cells are accompanied by ionic changes. The development of techniques allowing the measurement of such ion activities has contributed substantially to our understanding of normal and abnormal cellular function. Digital video microscopy, confocal laser scanning microscopy, and more recently multiphoton microscopy have allowed the precise spatial analysis of intracellular ion activity at the subcellular level in addition to measurement of its concentration. It is well known that Ca2+ regulates numerous physiological cellular phenomena as a second messenger as well as triggering pathological events such as cell injury and death. A number of methods have been developed to measure intracellular Ca2+. In this review, we summarize the advantages and pitfalls of a variety of Ca2+ indicators used in both optical and nonoptical techniques employed for measuring intracellular Ca2+ concentration.

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Section: B Ultraviolet-wavelength Excitation Fluorescent Indicatorsmentioning

confidence: 99%

Section: Confocal Laser Scanning Microscopymentioning

confidence: 99%

Measurement of Intracellular Calcium

Takahashi

Camacho

Lechleiter

et al. 1999

Physiological Reviews

674

511

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“…As shown in Fig. 4(a), I for the first time visualized [Ca 2+ ] i dynamics in isolated Ca 2+ ‐tolerant cardiomyocytes loaded with indo‐1, prepared from the heart, with a fluorescence microscope under whole‐cell voltage‐clamp mode 28,29 . The wave propagates rapidly, at a constant velocity of approximately 100 µm/s.…”

Section: Arrhythmogenic Disturbance Of Intracellular Calcium Ion Dynamentioning

confidence: 99%

Arrhythmogenic substrates in myocardial infarct

Takamatsu

2008

Pathology International

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Life-threatening ventricular tachyarrhythmias are common clinical complications in ischemic heart diseases, especially infarcted heart. Although electrophysiological mechanisms have been extensively clarified for the genesis of arrhythmias in myocardial infarct, arrhythmogenic substrates in the infarct that eventually lead to electrical derangements are not fully understood. This review focuses on the intracellular calcium ion (Ca2+) dynamics and connexin43 (Cx43) gap junctions that play pivotal roles in excitation/contraction processes and intercellular communication, respectively, in heart muscle cells. Recent development of Ca2+-sensitive fluorescent dyes as well as microscopy imaging techniques has contributed substantially to a more precise understanding of spatiotemporal aspects in the intra- and inter-cellular dynamics of Ca2+ in cardiomyocytes. Ca2+ waves, heterogeneous wave-like elevations of the intracellular Ca2+ concentrations ([Ca2+](i)) that develop under [Ca2+](i)-overloaded conditions of the injured myocardium, play an essential role in arrhythmias, especially in triggered arrhythmias. Alteration of Cx43-mediated electrical coupling, that is, gap junction remodeling that arises at myocyte-myocyte and myocyte-myofibroblast interfaces, would also be an important substrate for arrhythmias, especially re-entrant tachyarrhythmias. Clarification of these substrates would provide not only deeper insights into the upstream events of life-threatening tachyarrhythmias in the infarcted heart but also bases for new therapeutic strategies for cardiovascular diseases.

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“…spectrum-shifting probes with good measuring character istics. To date, confocal measurement of Ca2+ has used either the dual emission probe indo-l excited by the 351-nm line of high-power argon ion laser [19] or non-ratiometric imaging of other probes including fluo-3 [20], cal cium green [21] or fura red [22], This limitation of avail able excitation wavelengths imposes certain constraints on confocal imaging. For example, using both photomet ric recording and conventional imaging it is possible to differentiate between subcellular Ca2+ compartments in some cell types by taking advantage of the ability of probes such as fluo-3 (cytosolic), mag-fura-2 (calcium stores) and rhod-2 (mitochondria) to partition between different intracellular stores using appropriate loading conditions.…”

Section: Ion-dependent Fluorescent Probesmentioning

confidence: 99%

Applications of Confocal and Fluorescence Microscopy

Pedley

1997

Digestion

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Fluorescence microscopy has become a powerful tool for both the localization of cellular components in fixed cells, using target-specific fluorescent probes and labeled antibodies, and the fluorescence imaging of ions in single living cells. Despite its markedly lower spatial resolution when compared with electron microscopy, the essentially non-invasive nature of light microscopy provides a unique tool for examining cell behaviour at the level of the single cell bringing new insight into both cellular heterogeneities and cell-cell interactions. Further developments in both fluorescent probes and instrumentation should provide even more powerful tools to probe the mechanisms of cell function, both in vitro and in vivo.

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Confocal calcium imaging with ultraviolet laser- scanning microscopy and indo-1

Cited by 17 publications

References 16 publications

Measurement of Intracellular Calcium

Measurement of Intracellular Calcium

Arrhythmogenic substrates in myocardial infarct

Applications of Confocal and Fluorescence Microscopy

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