Real Time in Situ Confocal Imaging of Calcium Wave in the Perfused Whole Heart of the Rat

Hama, Takemitsu; Takahashi, Akiyuki; Ichihara, Akira; Takamatsu, Tetsuro

doi:10.1016/s0898-6568(97)00136-8

Cited by 37 publications

(32 citation statements)

References 23 publications

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“…However, FCS sometimes contains nonspecific esterases and causes hydrolysis of the AM before entering the cell. In addition to the comparative ease of incorporating these dyes into cultured and/or isolated cells, ester loading methods have also allowed loading of indicators into whole organs (e.g., isolated whole heart) while maintaining physiological conditions (136,217,251,357,372). On the other hand, AM loading has various problems including compartmentalization and incomplete hydrolysis (see sect.…”

Section: A Ester Loadingmentioning

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

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668

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: A Ester Loadingmentioning

confidence: 99%

Section: Confocal Laser Scanning Microscopymentioning

confidence: 99%

Measurement of Intracellular Calcium

Takahashi

Camacho

Lechleiter

et al. 1999

Physiological Reviews

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668

511

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“…1,2 Confocal (1-photon) and 2-photon imaging modalities have been used to study calcium [3][4][5][6] and voltage [7][8][9][10] transients at cellular and subcellular 11 resolutions in the intact organ. Physiologically important parameters, such as sarcomere spacing 12,13 and intracellular calcium wave front speeds, [14][15][16][17] can also be directly measured. Given the anisotropic nature of myocardial tissue organization, quantification of measurements with a spatial component (distance or velocity) assumes that the cell orientation relative to the image plane is known or can be ignored.…”

mentioning

confidence: 99%

“…Rapid capture of cell orientation is especially relevant when considering measurements of calcium wave speed at the cell level in the dye-loaded intact heart, which is an increasingly popular experimental model. [14][15][16][17] Similar techniques may be applicable to diverse biological preparations that use optical sectioning microscopy to measure velocities. 54,55 We think that remote focusing microscopy techniques have the potential to lead to a better understanding of how function and structure are linked in the intact organ.…”

mentioning

confidence: 99%

Fast Measurement of Sarcomere Length and Cell Orientation in Langendorff-Perfused Hearts Using Remote Focusing Microscopy

Botcherby¹,

Corbett

Burton

et al. 2013

Circulation Research

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Objective: We developed a method for measuring SL and regional cell orientation using remote focusing microscopy, an emerging imaging modality that can capture light from arbitrary oblique planes within a sample. Methods and Results:We present a protocol that unambiguously and quickly determines cell orientation from user-selected areas in a field of view by imaging 2 oblique planes that share a common major axis with the cell. We demonstrate the effectiveness of the technique in establishing single-cell SL in Langendorff-perfused hearts loaded with the membrane dye di-4-ANEPPS. Conclusions:

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“…We developed a system for in situ imaging of [Ca 2+ ]i equipped with a multipinhole-type confocal scanning device, which enabled us to visualize real-time X-Y images of Ca 2+ waves [11,12]. Using this system on Langendorff-perfused rat hearts, we observed [Ca 2+ ] i dynamics in the in situ whole heart of a rat with real-time confocal single-photon excitation microscopy.…”

Section: Ca 2+ Imaging In a Whole Heart With Realtime Two-photon Ementioning

confidence: 99%

Real-Time Two-Photon Microscopy and Its Application for In Situ Imaging.

Kaneko

Fujita

Tanaka

et al. 2001

Acta Histochem. Cytochem

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The recent developments in fluorescentprobes and laser technologies have provided us opportunities to observe the biological functions in living organs and tissues under a microscope. Although single-photon excitation confocal microscopy is an indispensable tool for observing these specimens, the observable depth of this microscopy is still not enough to analyze the functions of whole parts of organs in living organisms. To observe the deeper parts, we have attempted to develop new real-time two-photon excitation microscopy equipped with a microlens-arrayed multipinhole scanning device. Using this real-time twophoton microscope, we examined the dynamics of the intracellular calcium concentration ([Ca 2+ ] i ) in the cardiac myocytes in the rat whole heart to understand their physiological functions and mechanisms. We could observe Ca 2+ waves on the surface in the whole heart. However, we still need higher-energy laser power for twophoton excitation and visualization of [Ca 2+ ] i dynamics in the deeper parts of the whole heart.

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Real Time in Situ Confocal Imaging of Calcium Wave in the Perfused Whole Heart of the Rat

Cited by 37 publications

References 23 publications

Measurement of Intracellular Calcium

Measurement of Intracellular Calcium

Fast Measurement of Sarcomere Length and Cell Orientation in Langendorff-Perfused Hearts Using Remote Focusing Microscopy

Real-Time Two-Photon Microscopy and Its Application for In Situ Imaging.

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