Name: imaging membrane potential with two types of genetically encoded fluorescent voltage sensors
Uploaded: 2016-02-04T14:00:00.000+00:00
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Description: Watch this Scientific Journal Video about Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors at JoVE.com

This work was supported by the World Class Institute (WCI) program of the National Research Foundation of Korea funded by Ministry of Education, Science, and Technology of Korea Grant WCI 2009-003 and Korea Institute of Science and Technology Institutional Program Project 2E24210. Sungmoo Lee was supported by Global Ph.D. Fellowship program (NRF-2013H1A2A1033344) of the National Research Foundation (NRF) under the Ministry of Education (MOE, Korea).

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著者らは、開示することは何もありません。

神経系は、いくつかの異なる方法で電圧を使用して、阻害は、シナプス入力がわずかな脱分極および比較的大きな電圧変化における活動電位の結果を引き起こし、若干の過分極を引き起こします。 GEVIsによって膜電位の変化を測定する能力は、同時に神経回路のいくつかの成分を分析の有望な可能性を提供しています。この報告書では、GEVIsを使用して、膜電位のイメージングの変更のため?...

この論文の主要な焦点は、遺伝的にコード化された蛍光タンパク質を用いたin vitroでの膜電位の変化の光学イメージングを実証することです。膜電位のイメージングの変化は、神経回路の活動を研究のエキサイティングな可能性を提供しています。蛍光強度の変化、膜電位の結果の変化は、カメラの各画素は、神経活動の非侵入測定を可能にする代理電極になったとき。 40年以上にわたり、有機電位感受性色素は、1-4膜電位の変化を観察するために有用でした。しかし、これらの染料は、細胞特異性を欠きます。また、いくつかの細胞型は染色が困難です。遺伝的にコードされた電圧インジケータ（GEVIs）蛍光電位感受性プローブ​​を発現特に検討される細胞を有することにより、これらの制限を克服します。 GEVIsの3つのクラスがあります。 GEVIの最初のクラスは、VOを使用しています単一の蛍光タンパク質（FP）5-9または蛍光共鳴エネルギー移動（FRET）ペア10-12のいずれかの電圧センシングホスファターゼからltage感知ドメイン。センサの第二のクラスは、蛍光指示薬直接13-15として、またはエレクトロクロミックFRET 16,17を介して微生物のロドプシンを使用しています。第三のクラスは、2つのコンポーネント、膜アンカーFPおよび膜結合消光染料18-20である第2の成分である遺伝的要素を利用しています。第二および第三のクラスは 、in vitroおよびスライス実験19,20 内のために有用であるが、センサーの唯一の最初のクラスは現在、in vivoでの 6を分析するのに有用です。 この報告では 、in vitroでGEVIsの最初のクラス（ 図1）を使用して、膜電位のイメージングを実証します。電圧センサのこの最初のクラスは、in vivoイメージングに移行するのが最も簡単です。 GEVIs U以来FPに融合された電圧センシングドメインは約センサのロドプシンクラスより明るい50倍されるtilizing、それらはむしろ非常に強力なレーザを必要とするよりも、アーク灯照明を用いて撮像することができます。明るさの格差の別の結果は、GEVIsの最初のクラスを簡単に脳の自家蛍光を超えることができるということです。ロドプシンベースのプローブはできません。センサの第三のクラスは、ファーストクラスと同じくらい明るいですが、in vivoで投与することが困難である化学消光剤の添加を必要とします。 我々は、したがって、単一のFP（Bongwoori）8とFRETペア（ナビ2）12からなるプローブとプローブの取得のデモンストレーションを行います。ナビ2.242とナビ2.244という名前の緑色蛍光ドナー、クローバーから成る11、および赤色蛍光アクセプター、mRuby2、 - VSFP-CR（クローバー-mRuby2電位感受性蛍光タンパク質）の蝶のバージョンでは、FRETがこのレポートで構築されています<s&gt; 12アップ。録音のこれらのタイプの導入は、研究者に提供できる情報GEVIsの種類のより良い理解を与える必要があります。 <img alt="図1" src="/files/ftp_upload/53566/53566fig1.jpg" /> 図1.この報告書 （A） で撮像された遺伝的にコードされた電圧インジケータ（GEVIs）膜貫通電圧センシングドメインおよび蛍光タンパク質を持つGEVIベースのモノFPの二種類 。 （B）の膜貫通電圧センシングドメインで構成されるFRETベースGEVI、FRETドナーとアクセプター。 <a href="https://www.jove.com/files/ftp_upload/53566/53566fig1large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a>

<table><tbody><tr><td>Inverted Microscope</td><td>Olympus</td><td>IX71</td><td></td></tr><tr><td>60X objective lens (numerical aperture = 1.35)</td><td>Olympus</td><td>UPLSAPO 60XO</td><td></td></tr><tr><td>Excitation filter</td><td>Semrock</td><td>&amp;nbsp;FF02-472/30</td><td>For voltage imaging of super ecliptic pHluorin in Bongwoori</td></tr><tr><td>Dichroic mirror</td><td>Semrock</td><td>FF495-Di03-25x36</td><td></td></tr><tr><td>Emission filter</td><td>Semrock</td><td>FF01-497/LP</td><td></td></tr><tr><td>75W Xenon arc lamp</td><td>CAIRN</td><td>OptoSource Illuminator</td><td>LEDs and lasers are also effective light sources</td></tr><tr><td>Slow speed CCD camera</td><td>Hitachi</td><td>KP-D20BU</td><td></td></tr><tr><td>Dual port camera adaptor</td><td>Olympus</td><td>U-DPCAD</td><td></td></tr><tr><td>High speed CCD camera</td><td>RedShirtImaging, LLC</td><td>NeuroCCD-SM</td><td></td></tr><tr><td>Image splitter</td><td>CAIRN</td><td>Optosplit 2</td><td></td></tr><tr><td>Excitation filter</td><td>Semrock</td><td>FF01-475/23-25</td><td>For voltage imaging of FRET pair based GEVI consisting of Clover and mRuby2)</td></tr><tr><td>Dichroic mirror</td><td>Semrock</td><td>FF495-Di03-25x36</td><td></td></tr><tr><td>Emission filter</td><td>Chroma</td><td>ET520/40</td><td></td></tr><tr><td>Dichroic mirror</td><td>Semrock</td><td>FF560-FDi01-25X36</td><td></td></tr><tr><td>Emission filter</td><td>Chroma</td><td>ET645/75</td><td></td></tr><tr><td>Vibration isolation system</td><td>Kinetic systems</td><td>250BM-IC, 5702E-3036-31</td><td></td></tr><tr><td>Patching chamber</td><td>Warner instruments</td><td>RC-26G, 64-0235</td><td></td></tr><tr><td>#0 Micro Coverglass (22 x 40 mm)</td><td>Electron Microscopy Sciences</td><td>72198-20</td><td></td></tr><tr><td>Temperature controller</td><td>Warner instruments</td><td>TC-344B</td><td></td></tr><tr><td>#0 (0.08 ~ 0.13 mm) - 10 mm diameter glass coverslip</td><td>Ted Pella</td><td>260366</td><td></td></tr><tr><td>Lipofection agent</td><td>Life Technologies</td><td>11668-027</td><td></td></tr><tr><td>Calcium phosphate reagent</td><td>Clontech - Takara</td><td>631312</td><td></td></tr><tr><td>Patch clamp amplifier</td><td>HEKA</td><td>EPC 10 USB amplifier</td><td></td></tr><tr><td>Multi-channel data acquisition software</td><td>HEKA</td><td>Patchmaster</td><td></td></tr><tr><td>Image acquisition and analysis software</td><td>RedShirtImaging</td><td>Neuroplex</td><td></td></tr><tr><td>Spreadsheet application software</td><td>Microsoft</td><td>Microsoft Excel 2010</td><td></td></tr><tr><td>Data analysis software</td><td>OriginLab</td><td>OriginPro 8.6.0</td><td></td></tr><tr><td>Demagnifier</td><td>Qioptiq LINOS</td><td>Optem standard camera coupler 0.38x SC38 J clamp</td><td></td></tr><tr><td>Confocal microscope</td><td>Nikon</td><td>Nikon A1R confocal microscope</td><td></td></tr><tr><td>Anti-fade reagent</td><td>Life Technologies</td><td>P36930</td><td></td></tr></tbody></table>

imaging membrane potential with two types of genetically encoded fluorescent voltage sensors

Genetically encoded voltage indicators (GEVIs) have improved to the point where they are beginning to be useful for in vivo recordings. While the ultimate goal is to image neuronal activity in vivo, one must be able to image activity of a single cell to ensure successful in vivo preparations. This procedure will describe how to image membrane potential in a single cell to provide a foundation to eventually image in vivo. Here we describe methods for imaging GEVIs consisting of a voltage-sensing domain fused to either a single fluorescent protein (FP) or two fluorescent proteins capable of F&#246;rster resonance energy transfer (FRET) in vitro. Using an image splitter enables the projection of images created by two different wavelengths onto the same charge-coupled device (CCD) camera simultaneously. The image splitter positions a second filter cube in the light path. This second filter cube consists of a dichroic and two emission filters to separate the donor and acceptor fluorescent wavelengths depending on the FPs of the GEVI. This setup enables the simultaneous recording of both the acceptor and donor fluorescent partners while the membrane potential is manipulated via whole cell patch clamp configuration. When using a GEVI consisting of a single FP, the second filter cube can be removed allowing the mirrors in the image splitter to project a single image onto the CCD camera.

一過性にトランスフェクトされた細胞は、有意な蛍光強度の変化と原形質膜発現の程度を示すことができます。でも同じカバースリップ上でいくつかのセルは、内部蛍光のレベルを変化させる必要があります。これは、細胞によって吸収されるトランスフェクション剤の使用量が最も可能性が高いです。時折、あまりにも多くの式は（明るい、高い内部蛍光で、細胞?...

Watch this Scientific Journal Video about Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors at JoVE.com

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

遺伝的にコードされた蛍光電圧センサーの2種類のイメージング膜電位

A method for imaging changes in membrane potential using genetically encoded voltage indicators is described.

Genetically encoded voltage indicators (GEVIs) have improved to the point where they are beginning to be useful for in vivo recordings. While the ultimate ...

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Neuroscience

10.7K ビュー.  Seoul National University. A method for imaging changes in membrane potential using genetically encoded voltage indicators is described. 

ビデオ: Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

Biosensing Motor Neuron Membrane Potential in Live Zebrafish Embryos

The protocols described here are designed to allow researchers to study cell communication without altering the integrity of the environment in which the cells are located. Specifically, they have been developed to analyze the electrical activity of excitable cells, such as spinal neurons. In such a scenario, it is crucial to preserve the integrity of the spinal cell, but it is also important to preserve the anatomy and physiological shape of the systems involved. Indeed, the comprehension of the manner in which the nervous system-and other complex systems-works must be based on a systemic approach. For this reason, the live zebrafish embryo was chosen as a model system, and the spinal neuron membrane voltage changes were evaluated without interfering with the physiological conditions of the embryos. 
Here, an approach combining the employment of zebrafish embryos with a FRET-based biosensor is described. Zebrafish embryos are characterized by a very simplified nervous system and are particularly suited for imaging applications thanks to their transparency, allowing for the employment of fluorescence-based voltage indicators at the plasma membrane during zebrafish development. The synergy between these two components makes it possible to analyze the electrical activity of the cells in intact living organisms, without perturbing the physiological state. Finally, this non-invasive approach can co-exist with other analyses (e.g., spontaneous movement recordings, as shown here).

Protocols described here allow for the study of the electrical properties of excitable cells in the most non-invasive physiological conditions by employing zebrafish embryos in an in vivo system together with a fluorescence resonance energy transfer (FRET)-based genetically encoded voltage indicator (GEVI) selectively expressed in the cell type of interest.

生きたゼブラフィッシュ胚における運動ニューロン膜電位のバイオセンシング

The protocols described here are designed to allow researchers to study cell communication without altering the integrity of the environment in which the ...

発生生物学

Voltage-sensitive Dye Recording from Axons, Dendrites and Dendritic Spines of Individual Neurons in Brain Slices

Understanding the biophysical properties and functional organization of single neurons and how they process information is fundamental for understanding how the brain works. The primary function of any nerve cell is to process electrical signals, usually from multiple sources. Electrical properties of neuronal processes are extraordinarily complex, dynamic, and, in the general case, impossible to predict in the absence of detailed measurements. To obtain such a measurement one would, ideally, like to be able to monitor, at multiple sites, subthreshold events as they travel from the sites of origin on neuronal processes and summate at particular locations to influence action potential initiation. This goal has not been achieved in any neuron due to technical limitations of measurements that employ electrodes. To overcome this drawback, it is highly desirable to complement the patch-electrode approach with imaging techniques that permit extensive parallel recordings from all parts of a neuron. Here, we describe such a technique - optical recording of membrane potential transients with organic voltage-sensitive dyes (Vm-imaging) - characterized by sub-millisecond and sub-micrometer resolution. Our method is based on pioneering work on voltage-sensitive molecular probes 2. Many aspects of the initial technology have been continuously improved over several decades 3, 5, 11. Additionally, previous work documented two essential characteristics of Vm-imaging. Firstly, fluorescence signals are linearly proportional to membrane potential over the entire physiological range (-100 mV to +100 mV; 10, 14, 16). Secondly, loading neurons with the voltage-sensitive dye used here (JPW 3028) does not have detectable pharmacological effects. The recorded broadening of the spike during dye loading is completely reversible 4, 7. Additionally, experimental evidence shows that it is possible to obtain a significant number (up to hundreds) of recordings prior to any detectable phototoxic effects 4, 6, 12, 13. At present, we take advantage of the superb brightness and stability of a laser light source at near-optimal wavelength to maximize the sensitivity of the Vm-imaging technique. The current sensitivity permits multiple site optical recordings of Vm transients from all parts of a neuron, including axons and axon collaterals, terminal dendritic branches, and individual dendritic spines. The acquired information on signal interactions can be analyzed quantitatively as well as directly visualized in the form of a movie.

An imaging technique for monitoring of membrane potential changes with sub-micrometer spatial and sub-millisecond temporal resolution is described. The technique, based on laser excitation of voltage-sensitive dyes, allows measurements of signals in axons and axon collaterals, terminal dendritic branches, and individual dendritic spines.

軸索、樹状突起および脳スライスにおける個々のニューロンの樹状突起棘から電位感受性色素記録

Understanding the biophysical properties and functional organization of single neurons and how they process information is fundamental for understanding ...

神経科学

Measuring the Induced Membrane Voltage with Di-8-ANEPPS

Placement of a cell into an external electric field causes a local charge redistribution inside and outside of the cell in the vicinity of the cell membrane, resulting in a voltage across the membrane. This voltage, termed the induced membrane voltage (also induced transmembrane voltage, or induced transmembrane potential difference) and denoted by &Delta;&Phi;, exists only as long as the external field is present. If the resting voltage is present on the membrane, the induced voltage superimposes (adds) onto it. By using one of the potentiometric fluorescent dyes, such as di-8-ANEPPS, it is possible to observe the variations of &Delta;&Phi; on the cell membrane and to measure its value noninvasively. di-8-ANEPPS becomes strongly fluorescent when bound to the lipid bilayer of the cell membrane, with the change of the fluorescence intensity proportional to the change of &Delta;&Phi;. This video shows the protocol for measuring &Delta;&Phi; using di-8-ANEPPS and also demonstrates the influence of cell shape on the amplitude and spatial distribution of &Delta;&Phi;.

External electric field induces a voltage on the membrane of a cell, termed the induced membrane voltage (&Delta;&Phi;). By using the potentiometric dye di-8-ANEPPS, it is possible to measure the &Delta;&Phi; noninvasively. This video shows the protocol for measuring &Delta;&Phi; using di-8-ANEPPS.

ジ- 8 - ANEPPSで誘起膜電位の測定

Placement of a cell into an external electric field causes a local charge redistribution inside and outside of the cell in the vicinity of the cell ...

生物学

Optical Imaging of Isolated Murine Ventricular Myocytes

The ability to isolate adult cardiac myocytes has permitted researchers to study a variety of cardiac pathologies at the single cell level. While advances in calcium sensitive dyes have permitted the robust optical recording of single cell calcium dynamics, recording of robust transmembrane optical voltage signals has remained difficult. Arguably, this is because of the low single to noise ratio, phototoxicity, and photobleaching of traditional potentiometric dyes. Therefore, single cell voltage measurements have long been confined to the patch clamp technique which while the gold standard, is technically demanding and low throughput. However, with the development of novel potentiometric dyes, large, fast optical responses to changes in voltage can be obtained with little to no phototoxicity and photobleaching. This protocol describes in detail how to isolate adult murine myocytes which can be used for cellular shortening, calcium, and optical voltage measurements. Specifically, the protocol describes how to use a ratiometric calcium dye, a single-excitation calcium dye, and a single excitation voltage dye. This approach can be used to assess the cardiotoxicity and arrhythmogenicity of various chemical agents. While phototoxicity is still an issue at the single cell level, methodology is discussed on how to reduce it.

We present the methodology for the isolation of murine myocytes and how to obtain voltage or calcium traces simultaneously with sarcomere shortening traces using fluorescence photometry with simultaneous digital cell geometry measurements.

単離マウス心室筋細胞の光学イメージング

The ability to isolate adult cardiac myocytes has permitted researchers to study a variety of cardiac pathologies at the single cell level. While advances ...

生物工学

Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Conventional intracellular microelectrode techniques to quantify cardiomyocyte electrophysiology are extremely complex, labor intensive, and typically carried out in low throughput. Rapid and ongoing expansion of induced pluripotent stem cell (iPSC) technology presents a new standard in cardiovascular research and alternate methods are now necessary to increase throughput of electrophysiological data at a single cell level. VF2.1Cl is a recently derived voltage sensitive dye which provides a rapid single channel, high magnitude response to fluctuations in membrane potential. It possesses kinetics superior to those of other existing voltage indicators and makes available functional data equivalent to that of traditional microelectrode techniques. Here, we demonstrate simplified, non-invasive action potential characterization in externally paced human iPSC derived cardiomyocytes using a modular and highly affordable photometry system.

Here we describe optical acquisition and characterization of action potentials from induced pluripotent stem cell derived cardiomyocytes using a high-speed modular photometry system.

ヒト人工多能性幹細胞由来心筋細胞における単細胞光学作用電位測定

Conventional intracellular microelectrode techniques to quantify cardiomyocyte electrophysiology are extremely complex, labor intensive, and typically ...

Ex Vivo Imaging of Cell-specific Calcium Signaling at the Tripartite Synapse of the Mouse Diaphragm

The electrical activity of cells in tissues can be monitored by electrophysiological techniques, but these are usually limited to the analysis of individual cells. Since an increase of intracellular calcium (Ca2+) in the cytosol often occurs because of the electrical activity, or in response to a myriad of other stimuli, this process can be monitored by the imaging of cells loaded with fluorescent calcium-sensitive dyes.&#160; However, it is difficult to image this response in an individual cell type within whole tissue because these dyes are taken up by all cell types within the tissue. In contrast, genetically encoded calcium indicators (GECIs) can be expressed by an individual cell type and fluoresce in response to an increase of intracellular Ca2+, thus permitting the imaging of Ca2+ signaling in entire populations of individual cell types. Here, we apply the use of the GECIs GCaMP3/6 to the mouse neuromuscular junction, a tripartite synapse between motor neurons, skeletal muscle, and terminal/perisynaptic Schwann cells. We demonstrate the utility of this technique in classic ex vivo tissue preparations. Using an optical splitter, we perform dual-wavelength imaging of dynamic Ca2+ signals and a static label of the neuromuscular junction (NMJ) in an approach that could be easily adapted to monitor two cell-specific GECI or genetically encoded voltage indicators (GEVI) simultaneously. Finally, we discuss the routines used to capture spatial maps of fluorescence intensity. Together, these optical, transgenic, and analytic techniques can be employed to study the biological activity of distinct cell subpopulations at the NMJ in a wide variety of contexts.

Ex Vivo Imaging of Cell-specific Calcium Signaling at the Tripartite Synapse of the Mouse Diaphragm

Here we present a protocol to image calcium signaling in populations of individual cell types at the murine neuromuscular junction.

Ex Vivoセル固有カルシウム マウス ダイヤフラムの三者のシナプスでシグナル伝達のイメージング

The electrical activity of cells in tissues can be monitored by electrophysiological techniques, but these are usually limited to the analysis of ...

<meta charset="utf8"></head><body>遺伝子改変スパイキングHEK細胞に対するエレクトロポレーション効果の評価

Monitoring Electroporation-Induced Changes in Action Potential Generation in Genetically Engineered Tet-On Spiking HEK cells

Excitable cells such as&#160;neuronal and muscle cells can be primary targets in rapidly emerging electroporation-based treatments. However, they can be affected by electric pulses even in therapies where they are not the primary targets, and this can cause adverse side effects. Therefore, to optimize the electroporation-based treatments of excitable and non-excitable tissues, there is a need to study the effects of electric pulses on excitable cells, their ion channels, and excitability in vitro. For this purpose, a protocol was developed for optical monitoring of changes in action potential generation due to electroporation on a simple excitable cell model of genetically engineered tet-on spiking HEK cells. With the use of a fluorescent potentiometric dye, the changes in transmembrane voltage were monitored under a fluorescence microscope, and relevant parameters of cell responses were extracted automatically with a MATLAB application. This way, the excitable cell responses to different electric pulses and the interplay between excitation and electroporation could&#160;be efficiently evaluated.

<meta charset="utf8"></head><body>遺伝子改変Tet-OnスパイキングHEK細胞におけるエレクトロポレーション誘発性活動電位生成の変化のモニタリング

For studying responses of excitable cells in vitro, the protocol describes optical monitoring of changes in action potential generation due to electroporation on a simple excitable cell model of genetically engineered tet-on spiking HEK cells as well as changes in transmembrane voltage with automated extraction of relevant parameters.

遺伝子改変Tet-OnスパイキングHEK細胞におけるエレクトロポレーション誘発性活動電位生成の変化のモニタリング

Excitable cells such as&#160;neuronal and muscle cells can be primary targets in rapidly emerging electroporation-based treatments. However, they can be ...

Real-Time Fluorescent Measurement of Synaptic Functions in Models of Amyotrophic Lateral Sclerosis

Before neuronal degeneration, the cause of motor and cognitive deficits in patients with amyotrophic lateral sclerosis (ALS) and/or frontotemporal lobe dementia (FTLD) is dysfunction of communication between neurons and motor neurons and muscle. The underlying process of synaptic transmission involves membrane depolarization-dependent synaptic vesicle fusion and the release of neurotransmitters into the synapse. This process occurs through localized calcium influx into the presynaptic terminals where synaptic vesicles reside. Here, the protocol describes fluorescence-based live-imaging methodologies that reliably report depolarization-mediated synaptic vesicle exocytosis and presynaptic terminal calcium influx dynamics in cultured neurons.
Using a styryl dye that is incorporated into synaptic vesicle membranes, the synaptic vesicle release is elucidated. On the other hand, to study calcium entry, Gcamp6m is used, a genetically encoded fluorescent reporter. We employ high potassium chloride-mediated depolarization to mimic neuronal activity. To quantify synaptic vesicle exocytosis unambiguously, we measure the loss of normalized styryl dye fluorescence as a function of time. Under similar stimulation conditions, in the case of calcium influx, Gcamp6m fluorescence increases. Normalization and quantification of this fluorescence change are performed in a similar manner to the styryl dye protocol. These methods can be multiplexed with transfection-based overexpression of fluorescently tagged mutant proteins. These protocols have been extensively used to study synaptic dysfunction in models of FUS-ALS and C9ORF72-ALS, utilizing primary rodent cortical and motor neurons. These protocols easily allow for rapid screening of compounds that may improve neuronal communication. As such, these methods are valuable not only for the study of ALS but for all areas of neurodegenerative and developmental neuroscience research.

Two related methods are described to visualize subcellular events required for synaptic transmission. These protocols enable the real-time monitoring of the dynamics of presynaptic calcium influx and synaptic vesicle membrane fusion using live-cell imaging of in vitro cultured neurons.

筋萎縮性側索硬化症のモデルにおけるシナプス機能のリアルタイム蛍光測定

Before neuronal degeneration, the cause of motor and cognitive deficits in patients with amyotrophic lateral sclerosis (ALS) and/or frontotemporal lobe ...

遺伝的にコードされた蛍光電圧センサーの2種類のイメージング膜電位

要約

さらに動画を探す

この動画の章

生きたゼブラフィッシュ胚における運動ニューロン膜電位のバイオセンシング

軸索、樹状突起および脳スライスにおける個々のニューロンの樹状突起棘から電位感受性色素記録

ジ- 8 - ANEPPSで誘起膜電位の測定

単離マウス心室筋細胞の光学イメージング

ヒト人工多能性幹細胞由来心筋細胞における単細胞光学作用電位測定

Ex Vivoセル固有カルシウムマウスダイヤフラムの三者のシナプスでシグナル伝達のイメージング

遺伝子改変Tet-OnスパイキングHEK細胞におけるエレクトロポレーション誘発性活動電位生成の変化のモニタリング

遺伝子改変Tet-OnスパイキングHEK細胞におけるエレクトロポレーション誘発性活動電位生成の変化のモニタリング

遺伝子改変Tet-OnスパイキングHEK細胞におけるエレクトロポレーション誘発性活動電位生成の変化のモニタリング

筋萎縮性側索硬化症のモデルにおけるシナプス機能のリアルタイム蛍光測定

遺伝的にコードされた蛍光電圧センサーの2種類のイメージング膜電位

要約

さらに動画を探す

この動画の章

生きたゼブラフィッシュ胚における運動ニューロン膜電位のバイオセンシング

軸索、樹状突起および脳スライスにおける個々のニューロンの樹状突起棘から電位感受性色素記録

ジ- 8 - ANEPPSで誘起膜電位の測定

単離マウス心室筋細胞の光学イメージング

ヒト人工多能性幹細胞由来心筋細胞における単細胞光学作用電位測定

Ex Vivoセル固有カルシウム マウス ダイヤフラムの三者のシナプスでシグナル伝達のイメージング

遺伝子改変Tet-OnスパイキングHEK細胞におけるエレクトロポレーション誘発性活動電位生成の変化のモニタリング

遺伝子改変Tet-OnスパイキングHEK細胞におけるエレクトロポレーション誘発性活動電位生成の変化のモニタリング

遺伝子改変Tet-OnスパイキングHEK細胞におけるエレクトロポレーション誘発性活動電位生成の変化のモニタリング

筋萎縮性側索硬化症のモデルにおけるシナプス機能のリアルタイム蛍光測定

Ex Vivoセル固有カルシウムマウスダイヤフラムの三者のシナプスでシグナル伝達のイメージング