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FLEXcyte 96
FLEXcyte 96 -生理状态下记录细胞收缩
FLEXcyte 96带来了全新的心脏细胞的体外收缩力测量技术。该系统可利用干细胞分化的心肌细胞以更高的通量形式研究药理化合物的作用。 在96孔板中,单层细胞粘附在柔性基底上,并记录细胞层的同步收缩性。 这种独特的技术允许在接近心脏机械条件的原生环境中进行记录。 FLEXcyte 96是到手可用的系统,用于心脏毒性,安全性和功效筛选以及基础研究中的结构和功能收缩力记录。
FLEXcyte 96是 CardioExcyte 96的一个可选附件。FLEXcyte 96采用的记录板上的聚二甲基硅氧烷(PDMS)膜厚度小于10μm,表面经过精密修饰,具有天然人体心脏组织的生理弹性和强大的机械支撑。 当被培养基的重量偏转时,心肌细胞的节律性收缩使96孔中的膜向上提升。 通过测量偏转的变化,可以计算出机械应力。
该系统已经在许多干细胞分化的心肌细胞(Ncardia,FUJIFILM Cellular Dynamics, Inc. (FCDI),Axol和NEXEL)上进行了验证。 人源诱导多能干细胞分化的心肌细胞(hiPSC-CM)显示出正性肌力反应,不需要额外刺激“细胞成熟”。 此外还可以在患者来源的细胞上测量心脏收缩力。
FLEXcyte 96的记录不会遗漏细胞毒性反应的细微差别,可以可靠地测量心肌细胞的收缩力,以便在慢性和急性时间框架内进行安全性,有效性和毒性研究。 FLEXcyte 96是一种全自动设备,可并行记录96孔细胞。
FLEXcyte 96 的主要特征有:
CardioExcyte 96 / FLEXcyte 96系统包括一个小型的培养设备,使系统可独立于培养箱使用。 该设备放置在工作台顶部,同时保持对温度,气体混合和湿度的控制。该系统有出色的数据处理和输出软件包促进了效率和易用性 - 这是CardioExcyte 96 / FLEXcyte 96系统的重要组成部分。
CardioExcyte 96/FLEXcyte 96包装包含Integra Viaflo Assist,可提供自动多通道移液,用于细胞和化合物的可复制96孔板处理。
FLEXcyte 96包装包括一个培养系统。这意味着测量是在细胞培养箱外进行的,并且整个装置安装在实验室工作台上。FLEXcyte 96培养系统控制温度、气体混合和湿度。它适用于各种实验条件(如缺氧)。
FLEXcyte 96 SOL (Add-On)
平行进行光学刺激和收缩力研究:
刺激光学盖CardioExcyte 96/ FLEXcyte SOL使用led对转染了光门控离子通道(如Channelrhodopsin2 (ChR2))的细胞进行空间均匀刺激。用ChR2转染iPS心肌细胞后,细胞可被蓝光刺激。每一个光脉冲诱导细胞去极化,从而触发一个动作电位。光刺激优于电刺激的优点包括高度精确的时间,跳动网络的所有细胞都在光刺激的时间被刺激。相反,电刺激从电极传播到井的另一端,因此细胞受到渐进的刺激。精确计时节拍的平均节拍计算能够获得深入的化合物分析和浓度响应相关性。这是每个CardioExcyte 96/ FLEXcyte 96系统提供的CardioExcyte Control/ DataControl软件包的关键软件功能之一。
CardioExcyte 96/ FLEXcyte 96 Software package
自适应信号检测算法提取跳动事件的位置和值。除了拍间隔、振幅、上升和下降时间外,还检测脉冲宽度。此外,还识别和量化了搏动和心律失常(黄色)下的区域。
CardioExcyte/ FLEXcyte Control软件支持收缩性参数的在线分析。独特的平均节拍函数自动可视化从一个井的节拍轨迹的平均值,由标准偏差包围,表明节拍模式在记录井内的一致性,即蜂窝网络是否同步跳动。控制软件直观易用,同时在实验设置和记录数据的在线可视化方面非常强大。DataControl 96软件包是一个独立的软件包,用于加载和分析记录在CardioExcyte 96或FLEXcyte 96上的数据。可以保存和重新加载分析模板。集成了分析结果的自定义导出格式。
FLX-96
FLX-96 FLEXcyte膜板采用标准的96孔格式,允许自动液体处理和轻松集成到您的实验室程序中。经过7天的培养,细胞已经形成了生物杂交材料与膜和准备试验。为了在FLEXcyte 96板提供的生理环境中进一步成熟,细胞可以培养4周。FLX-96平板是innoVitro公司为FLEXcyte 96开发的专利创新产品。它在innoVitro总部内部生产和质量保证,并发货给我们的国际客户。FLEXcyte 96的聚二甲基硅氧烷(PDMS)膜厚度小于10 μm,经过复杂的表面修饰,结合了人体心脏组织的生理弹性和强大的机械支持。
技术原理:
胞外场电位是细胞产生的电位,例如神经或肌肉细胞的细胞外。 电生理学研究使用细胞外微电极研究这些电位。 在这些实验中,细胞外场电位被检测为电势。对于个体细胞,细胞外电位的时间过程理论上与跨膜电流成反比。
电阻抗是在施加电压时对电流的电阻的测量,具有幅度和相位。换句话说,它是在特定频率ω下单个复指数的电压 - 电流比。可以以欧姆直接测量或显示阻抗。
实际上:接种到电阻抗芯片或板上的细胞覆盖电极,从而产生更高的可检测电阻。阻抗测定可以在一个频率或一定频率范围内进行。取决于施加的频率ω,电子穿过或穿过细胞,因此可以检测细胞形态和覆盖率的不同参数。电阻抗测量适合于在长期研究中应用毒理学物质或影响细胞-细胞或者细胞-基质接触的物质时检测心肌细胞的收缩运动,细胞生长和运动以及细胞形态的变化。
光遗传学:光刺激触发细胞中动作电位光遗传学是一种可用于控制可兴奋细胞的新技术:可在表达特定光敏离子通道的细胞上用具有毫秒级时间精度的光脉冲对细胞进行去极化,从而准确地触发动作的电势。光遗传学在心脏安全性研究方面提供了巨大潜力:影响心肌细胞收缩的化合物可能以依赖于使用的方式影响离子通道和受体,从而对不同的搏动率产生不同的影响。利用这种技术,可以简单地利用光脉冲在不同跳动范围内使心肌细胞起搏,以发现毒素的使用依赖效应。在iPSC分化的心肌细胞中,光遗传学致动器 通道视紫红质已被证明非常适合并被接受为模型。用于通道视紫红质的瞬时转染试剂盒可从iPSC细胞提供者获得,例如NCardia,用于在心脏安全性研究中使用光遗传学。
数据与应用:
Detection of cardiac arrhythmia
FLEXcyte 96 data and applications:
Cells were kindly provided by NEXEL.
Contractile behavior of NEXEL cells. DataControl software allows an overview on mean beats including standard deviations from all 96-wells. Thus, arrhythmia or rhythmic beating can be judged at a glance.
Digoxin Concentration response curves
FLEXcyte 96 data and applications:
Cells were kindly provided by NEXEL.
Acute effects of digoxin on hiPSC cardiomyocytes (Cardiosight®-S). Overlay of mean beats at 60 min after application of several digoxin concentrations (1 nM – 10 µM). Bar graphs depict development of beating amplitude over 60 min for the same digoxin concentrations. Concentrations > 1 µM predominately promote cytotoxic effects.
FLEXcyte 96 data and applications:
Cells were kindly provided by FUJIFILM Cellular Dynamics.
Isoproterenol is enhancing contractile force. Upon application of increasing Isoproterenol concentrations a beating frequency increase is observed, along with an increased beating amplitude (positive force-frequency relationship).
Isoproterenol increases contraction force
FLEXcyte 96 data and applications:
Cells were kindly provided by FUJIFILM Cellular Dynamics.
Amplitude increase of contraction signals as % of control plotted against increasing Isoproterenol concentrations.
Kinase inhibitor A-674563 – Arrhythmogenic effects on cardiomyocytes
FLEXcyte 96 data and applications:
Cells were kindly provided by FUJIFILM Cellular Dynamics.
Exemplary arrhythmia as observed 24 h after application of Kinase inhibitors A-674563. Aftercontractions (regular intervals and irregular intervals) were observed.
Kinase inhibitor Gefitinib – Arrhythmogenic effects on cardiomyocytes
FLEXcyte 96 data and applications:
Cells were kindly provided by FUJIFILM Cellular Dynamics.
Exemplary arrhythmia as observed 24 h after application of Kinase inhibitor Gefitinib. Aftercontractions (regular intervals and irregular intervals) and Fibrillations were observed, respectively.
Kinase inhibitor Nilotinib – Arrhythmogenic effects on cardiomyocytes
FLEXcyte 96 data and applications:
Cells were kindly provided by FUJIFILM Cellular Dynamics.
Exemplary arrhythmia as observed 24 h after application of Kinase inhibitor Nilotinib. Aftercontractions (regular intervals and irregular intervals) and beat rate variability were observed.
Kinase inhibitors – Chronic compound effects on cardiomyocytes
FLEXcyte 96 data and applications:
Cells were kindly provided by FUJIFILM Cellular Dynamics.
Here Structural and/or functional toxicity of Kinase inhibitors Trametinib, Sorafenib, Nilotinib and A-674563 was investigated. Compound incubation time was 5 days.
During investigations of chronic effects it could take days and weeks until onset of adverse effects. The CardioExcyte and FLEXcyte technology and the consumables are perfectly suited for such studies.
Positive Inotropes - Omecamtiv Mecarbil myosin complex activation
FLEXcyte 96 data and applications:
Cells were kindly provided by NEXEL.
Omecamtiv mecarbil increased both the contraction amplitude as well as the duration of the contraction-relaxation-cycle, indicating the positive inotropic effect based on activation of the myosin complex. The symmetry of the contraction-relaxation-cycle was not changed.
Read the application note here.
发表文献:
2022 - Conventional rigid 2D substrates cause complex contractile signals in monolayers of human induced pluripotent stem cell-derived cardiomyocytes
2020 - Integration of mechanical conditioning into a high throughput contractility assay for cardiac safety assessment
2019 - Higher throughput in vitro cardiac contractility recordings under physiological mechanical conditions
2016 - Mechano-Pharmacological Characterization of Cardiomyocytes Derived from Human Induced Pluripotent Stem Cells
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资料下载:
CE 96实时无标记细胞功能测试仪.pdf 附件下载 (下载 0 次)
Nanion_Poster_SURFE2R_96SE_FLEXcyte96_2021_BPS_NCXCIPA (1)_1_1_translate.pdf 附件下载 (下载 0 次)
Nanion_CE96细胞监测.pdf 附件下载 (下载 1 次)
Nanion_Product_Flyer_CE_FL_Plates.pdf 附件下载 (下载 1 次)
心肌细胞应用专题.pdf 附件下载 (下载 0 次)
Nanion_FLEXcyte96产品彩页.pdf 附件下载 (下载 0 次)
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