Di-4-ANEPPS causes photodynamic damage to isolated cardiomyocytes

Schäffer, Péter; Ahammer, Helmut; Müller, Wolfram; Koidl, Bernd; Windisch, H.

doi:10.1007/bf00378533

Cited by 90 publications

(68 citation statements)

References 14 publications

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“…3, B and C). In contrast, adult ventricular myocytes loaded with di-4-ANEPPS or di-4-ANBDQBS developed marked prolongation in APD 90 during the second cycle of laser illumination (13,16). Taken together, these data indicate that optical AP recordings in di-4-ANB-DQBS-labeled human iPSC-CMs are stable over time and are relatively resistant to photodynamic damage.…”

Section: Stability Of Di-4-anbdqbs Optical Ap Recordings and Photodynmentioning

confidence: 77%

“…3A), nor did measurements of APD 10 , APD 50 , or spontaneous CL (data not shown). Next, we assessed the susceptibility to photodynamic damage using an intense laser illumination protocol that has been previously reported to perturb optical APs recordings in isolated adult ventricular myocytes loaded with either di-4-ANEPPS or di-4-ANBDQBS, respectively (13,16). Human iPSC-CMs loaded with di-4-ANBDQBS were sequentially exposed to three cycles of 2.5 min of continuous laser illumination interspersed by brief periods of recovery.…”

Section: Stability Of Di-4-anbdqbs Optical Ap Recordings and Photodynmentioning

confidence: 99%

“…P Ͼ 0.05 by repeated-measures ANOVA. B and C: human iPSC-CMs were exposed to an intense laser illumination protocol that has been previously reported to perturb optical APs recordings in adult ventricular myocytes loaded with either di-4-ANEPPS or di-4-ANBDQBS, respectively (13,16). Shaded rectangles denote time periods of laser exposure.…”

Section: Stability Of Di-4-anbdqbs Optical Ap Recordings and Photodynmentioning

confidence: 99%

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A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes

López-Izquierdo

Warren

Riedel

et al. 2014

American Journal of Physiology-Heart and Circulatory Physiology

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-Human induced pluripotent stem cell-derived cardiomyocyte (iPSC-CM)-based assays are emerging as a promising tool for the in vitro preclinical screening of QT interval-prolonging side effects of drugs in development. A major impediment to the widespread use of human iPSC-CM assays is the low throughput of the currently available electrophysiological tools. To test the precision and applicability of the near-infrared fluorescent voltage-sensitive dye 1-(4-sulfanatobutyl)-4-{␤[2-(di-n-butylamino)-6-naphthyl]butadienyl}quinolinium betaine (di-4-ANBDQBS) for moderate-throughput electrophysiological analyses, we compared simultaneous transmembrane voltage and optical action potential (AP) recordings in human iPSC-CM loaded with di-4-ANBDQBS. Optical AP recordings tracked transmembrane voltage with high precision, generating nearly identical values for AP duration (AP durations at 10%, 50%, and 90% repolarization). Human iPSC-CMs tolerated repeated laser exposure, with stable optical AP parameters recorded over a 30-min study period. Optical AP recordings appropriately tracked changes in repolarization induced by pharmacological manipulation. Finally, di-4-ANBDQBS allowed for moderate-throughput analyses, increasing throughput Ͼ10-fold over the traditional patchclamp technique. We conclude that the voltage-sensitive dye di-4-ANBDQBS allows for high-precision optical AP measurements that markedly increase the throughput for electrophysiological characterization of human iPSC-CMs. optical action potential; preclinical drug screening; QT prolongation; acquired long QT syndrome; stem cell model RECENT ADVANCES in induced pluripotent stem cell (iPSC) technology now allow for the routine derivation of patient-and disease-specific human iPSC cardiomyocyte (iPSC-CM) models of cardiovascular disease. The impact of this technology has far-reaching implications, ranging from drug discovery, preclinical drug screening, mechanistic understanding of disease processes, and approaches for personalized medicine. Human iPSC-CM-based assays are emerging as a promising tool for the in vitro preclinical screening of QT intervalprolonging side effects of drugs in development. A major impediment to the widespread use of human iPSC-CM assays is the low throughput of the currently available electrophysiological tools. The patch-clamp technique allows for direct measurement of transmembrane voltage (V m ) to precisely quantify the morphology and duration of atrial and ventricular action potentials (APs). However, the patch-clamp technique is a labor-intensive technique, which involves highly skilled, experienced individuals. Recently, the utility of a genetically encoded voltage-sensitive fluorescent reporter (ArcLight) was described in human embryonic stem cell-derived CMs (6). This reporter generated robust fluorescent signals from single cells, allowing for moderate-high throughput analyses, although the temporal response was delayed relative to directly measured V m . While voltage-sensitive dyes such as di-4-ANNEPS and di-8-ANNEPS allow ...

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Section: Stability Of Di-4-anbdqbs Optical Ap Recordings and Photodynmentioning

confidence: 77%

Section: Stability Of Di-4-anbdqbs Optical Ap Recordings and Photodynmentioning

confidence: 99%

Section: Stability Of Di-4-anbdqbs Optical Ap Recordings and Photodynmentioning

confidence: 99%

See 1 more Smart Citation

A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes

López-Izquierdo

Warren

Riedel

et al. 2014

American Journal of Physiology-Heart and Circulatory Physiology

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“…Pharmacological effects of the dye include phototoxicity 138 and increased contractility. 139 Yet, arguably, the major limitation of organ-level optical imaging is its restricted depth of penetration.…”

Section: Emerging Optical Imaging Modalitiesmentioning

confidence: 99%

Optical Imaging of the Heart

Efimov

Nikolski

Salama

2004

Circulation Research

369

296

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Abstract-Optical techniques have revolutionized the investigation of cardiac cellular physiology and advanced our understanding of basic mechanisms of electrical activity, calcium homeostasis, and metabolism. Although optical methods are widely accepted and have been at the forefront of scientific discoveries, they have been primarily applied at cellular and subcellular levels and considerably less to whole heart organ physiology. Numerous technical difficulties had to be overcome to dynamically map physiological processes in intact hearts by optical methods. Problems of contraction artifacts, cellular heterogeneities, spatial and temporal resolution, limitations of surface images, depth-offield, and need for large fields of view (ranging from 2ϫ2 mm 2 to 3ϫ3 cm 2 ) have all led to the development of new devices and optical probes to monitor physiological parameters in intact hearts. This review aims to provide a critical overview of current approaches, their contributions to the field of cardiac electrophysiology, and future directions of various optical imaging modalities as applied to cardiac physiology at organ and tissue levels. Key Words: optical mapping Ⅲ fluorescent probes Ⅲ electrophysiology Ⅲ arrhythmia Ⅲ defibrillation M ammalian physiology has an ingrained hierarchy with molecular and cellular physiology at its base, followed by the interactions of large populations of cells and organ systems, and finally the integration of multiple organ functions of an entire animal. For the past 4 decades, cardiovascular physiology has been dominated by a "reductionist" approach, focusing on cellular mechanisms. Major strides have been accomplished in our understanding of cellular mechanisms, including metabolism, intracellular signaling, trafficking, ion channel structure, function, and expression. With a greater understanding of cellular mechanisms came the growing realization that organs such as the heart are composed of several types of interacting cells with significant and important heterogeneities of properties, cell-to-cell coupling, and function within each group. Thus, an understanding of molecular and cellular mechanisms must still be integrated to explain the more complex organ system while taking into account spatial and temporal heterogeneities of cell functions throughout the organ.Unfortunately, experimental methodologies available for studies at the organ level are not as abundant as at the cellular scale. Nonoptical imaging modalities, including positron emission tomography, magnetic resonance, and ultrasound imaging have only started to bridge molecular and organ physiology using novel contrast agents. 1 On the other hand, optical modes of imaging, in combination with parametersensitive probes have already demonstrated their ability to overcome the problem of spatiotemporal resolution in two dimensions for a wide range of applications from single molecular events to in vivo whole animal physiology.Fluorescence has been used to measure a wide range of physiological parameters in cells and tissues...

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“…However, such an improvement may come at a price, causing photobleaching and phototoxicity. [11][12][13] The sensitivity of different dyes to increased light varies and should be determined for each dye individually.…”

Section: Introductionmentioning

confidence: 99%

Optical mapping at increased illumination intensities

et al. 2012

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Abstract. Voltage-sensitive fluorescent dyes have become a major tool in cardiac and neuro-electrophysiology. Achieving high signal-to-noise ratios requires increased illumination intensities, which may cause photobleaching and phototoxicity. The optimal range of illumination intensities varies for different dyes and must be evaluated individually. We evaluate two dyes: di-4-ANBDQBS (excitation 660 nm) and di-4-ANEPPS (excitation 532 nm) in the guinea pig heart. The light intensity varies from 0.1 to 5 mW∕mm 2 , with the upper limit at 5 to 10 times above values reported in the literature. The duration of illumination was 60 s, which in guinea pigs corresponds to 300 beats at a normal heart rate. Within the identified duration and intensity range, neither dye shows significant photobleaching or detectable phototoxic effects. However, light absorption at higher intensities causes noticeable tissue heating, which affects the electrophysiological parameters. The most pronounced effect is a shortening of the action potential duration, which, in the case of 532-nm excitation, can reach ∼30%. At 660-nm excitation, the effect is ∼10%. These findings may have important implications for the design of optical mapping protocols in biomedical applications.

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Di-4-ANEPPS causes photodynamic damage to isolated cardiomyocytes

Cited by 90 publications

References 14 publications

A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes

A near-infrared fluorescent voltage-sensitive dye allows for moderate-throughput electrophysiological analyses of human induced pluripotent stem cell-derived cardiomyocytes

Optical Imaging of the Heart

Optical mapping at increased illumination intensities

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