אנו רוצים להודות לגלעד זילברברג, מישל Pignatelli, תומאס ק ברגר, לוקה Gambazzi, וסוניה גרסיה לייעוץ רב ערך על שיפורים לאוטומציה הליך תיקון מהדק. אנו מודים Rajnish ראנג&#39;אן לקבלת ייעוץ וסיוע ביישום תוכנה יקרים. עבודה זו מומנה בחלקו על ידי האיחוד האירופי בפרויקט סינפסה ובחלקו על ידי התכנית למדע הגבולות האנושי.

<ol>
	<li>Perin, R., Berger, T. K., Markram, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+synaptic+organizing+principle+for+cortical+neuronal+groups.">A synaptic organizing principle for cortical neuronal groups.</a> Proc. Natl. Acad. Sci. U.S.A. 108, 5419-5424 (2011).</li><li>Berger, T. K., Silberberg, G., Perin, R., Markram, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brief+Bursts+Self-Inhibit+and+Correlate+the+Pyramidal+Network.">Brief Bursts Self-Inhibit and Correlate the Pyramidal Network.</a> PLoS Biol. 8, e1000473 (2010).</li><li>Fino, E., Yuste, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dense+inhibitory+connectivity+in+neocortex.">Dense inhibitory connectivity in neocortex.</a> Neuron. 69, 1188-1203 (2011).</li><li>Packer, A. M., Yuste, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dense,+Unspecific+Connectivity+of+Neocortical+Parvalbumin-Positive+Interneurons:+A+Canonical+Microcircuit+for+Inhibition.">Dense, Unspecific Connectivity of Neocortical Parvalbumin-Positive Interneurons: A Canonical Microcircuit for Inhibition.</a> J. Neurosci. 31, 13260-13271 (2011).</li><li>Berger, T. K., Perin, R., Silberberg, G., Markram, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Frequency-dependent+disynaptic+inhibition+in+the+pyramidal+network:+a+ubiquitous+pathway+in+the+developing+rat+neocortex.">Frequency-dependent disynaptic inhibition in the pyramidal network: a ubiquitous pathway in the developing rat neocortex.</a> J. Physiol. 587, 5411-5425 (2009).</li><li>Kodandaramaiah, S. B., Franzesi, G. T., Chow, B. Y., Boyden, E. S., Forest, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+whole-cell+patch-clamp+electrophysiology+of+neurons+in+vivo.">Automated whole-cell patch-clamp electrophysiology of neurons in vivo.</a> Nat. Methods. 9, 585-587 (2012).</li><li>Anastassiou, C. A., Perin, R., Markram, H., Koch, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ephaptic+coupling+of+cortical+neurons.">Ephaptic coupling of cortical neurons.</a> Nat. Neurosci. 14, 217-223 (2011).</li><li>Prakash, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two-photon+optogenetic+toolbox+for+fast+inhibition,+excitation+and+bistable+modulation.">Two-photon optogenetic toolbox for fast inhibition, excitation and bistable modulation.</a> Nat. Methods. 9, 1171-1179 (2012).</li><li>Papagiakoumou, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scanless+two-photon+excitation+of+channelrhodopsin-2.">Scanless two-photon excitation of channelrhodopsin-2.</a> Nat Methods. 7, 848-854 (2010).</li><li>Ko, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+specificity+of+local+synaptic+connections+in+neocortical+networks.">Functional specificity of local synaptic connections in neocortical networks.</a> Nature. 473, 87-91 (2011).</li><li>Wickersham, I. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monosynaptic+Restriction+of+Transsynaptic+Tracing+from+Single,+Genetically+Targeted+Neurons.">Monosynaptic Restriction of Transsynaptic Tracing from Single, Genetically Targeted Neurons.</a> Neuron. 53, 639-647 (2007).</li><li>Liang, C. W., Mohammadi, M., Santos, M. D., Tang, C. -. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterned+Photostimulation+with+Digital+Micromirror+Devices+to+Investigate+Dendritic+Integration+Across+Branch+Points.">Patterned Photostimulation with Digital Micromirror Devices to Investigate Dendritic Integration Across Branch Points.</a> J. Vis. Exp. (49), e2003 (2011).</li><li>Nikolenko, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SLM+Microscopy:+Scanless+Two-Photon+Imaging+and+Photostimulation+with+Spatial+Light+Modulators.">SLM Microscopy: Scanless Two-Photon Imaging and Photostimulation with Spatial Light Modulators.</a> Front Neural Circuits. 2, (2008).</li></ol>

החוקרים מצהירים כי אין להם אינטרסים כלכליים מתחרים.

שאלה מיידית שעולה בדרך כלל בדבר שיעור הצלחה של ההליך שתארנו מ. לשיעורי הצלחה גבוהים הכנה היא חיונית. טפטפות חייבת להיות פתחי קצה שנחשבים כמספיקים לאדם התאים שנרשם. סינון הפתרון תאיים, כדי למנוע טפטפות הסתומה הוא גם חשוב. טפטפות מאוד נקייה, טרי משכה הן דרישה אחרת. התפלג...

היכולת להקליט ולעורר מספר אתרים עם דיוק מיקרומטר היא מאוד שימושית עבור ניסוי השגת הבנה טובה יותר של מערכות עצביות. טכניקות רבות פותחו למטרה זו אבל אף מאפשרות הרזולוציה submillivolt מושגת על ידי טכניקת תיקון מהדק, חיונית ללימוד פעילות התת ופוטנציאלי postsynaptic בודדים. כאן אנו מכסים את פיתוחה של מערכת התיקון-clamp שנתי עשר האלקטרודה בעזרת מחשב שמטרתה הקלטה בו זמנית ומגרה מספר רב של תאים בודדים בדייקנות מספיק ללימוד קישוריות עצבית. אמנם יכולים להיות יזום יישומים רבים אחרים למערכת כזו, זה משאיל את עצמו במיוחד גם ללימוד קישוריות הסינפטית בהתחשב בעובדה שמספר חיבורים אפשריים בתוך קבוצה של נוירונים גדל באופן פרופורציונלי לריבוע מספר הנוירונים בשאלה. לכן, בזמן שמערכת עם שלוש אלקטרודות מאפשרת בדיקהמופע של עד שישה חיבורים ולרוב הקלטה אחת, הקלטת עשרה נוירונים מאפשר בדיקת ההתרחשות של עד 132 קשרים ולעתים קרובות התבוננות למעלה מתריסר (איור 1). התצפית של עשרות חיבורים בו זמנית מאפשרת לנתח את הארגון של רשתות קטנות ולהסיק מאפיינים סטטיסטיים של מבנה הרשת שלא יכול להיות נחקר אחר 1. יתר על כן, גירוי מדויק של תאים רבים גם מאפשר כימות של גיוס של תאי postsynaptic 2.

<table><tbody><tr><td>Microscope</td><td>Olympus</td><td>BX51WI</td><td>40X Immersion Objective</td></tr><tr><td>Manipulators</td><td>Luigs Neumann</td><td>SM-5</td><td>Serial protocol used</td></tr><tr><td>Amplifiers</td><td>Axon Instruments</td><td>MultiClamp 700B</td><td>SDK used</td></tr><tr><td>Camera</td><td>Till Photonics</td><td>VS 55</td><td>BNC analog output</td></tr><tr><td>Framegrabber</td><td>Data Translation</td><td>DT3120</td><td>SDK used</td></tr><tr><td>Oscilloscopes</td><td>Tektronix</td><td>TDS 2014</td><td>Serial communication</td></tr><tr><td>Data acquisition</td><td>InstruTECH</td><td>ITC 1600</td><td></td></tr><tr><td>Data acquisition</td><td>National Instruments</td><td>PCI-6221</td><td>Library used (.dll)</td></tr><tr><td>Pressure valve</td><td>SMC</td><td>SMC070C-6BG-32</td><td></td></tr><tr><td>Pressure sensor</td><td>Honeywell</td><td>24PCDFA6G</td><td></td></tr><tr><td>Membrane pump</td><td>Schego</td><td>Optimal</td><td></td></tr></tbody></table>

a computer-assisted multi-electrode patch-clamp system

טכניקת התיקון-clamp היא כיום השיטה הכי המבוססת היטב להקלטת פעילות חשמלית מנוירונים בודדים או תאי subcellular שלהם. עם זאת, להשגת הקלטות יציבה, גם מהתאים בודדים, נותר הליך מורכב למדי זמן רב. אוטומציה של שלבים רבים בשילוב עם תצוגת מידע יעילה יכולה מאוד לסייע experimentalists בביצוע מספר רב יותר של הקלטות עם אמינות גבוהה יותר ובפחות זמן. על מנת להשיג את הקלטות בקנה מידה גדולה שסכמנו את הגישה היעילה ביותר היא לא automatize באופן מלא בתהליך, אבל כדי לפשט את הצעדים הניסיוניים ולהפחית את הסיכוי לטעויות אנוש תוך שילוב ניסיונו של הנסיין ומשוב חזותי ביעילות. עם מטרות אלה בחשבון שפיתחנו מערכת ממוחשבת המרכזת את כל הבקרות הנדרשות לניסוי רב האלקטרודה תיקון מהדק בממשק יחיד, commercgamepad האלחוטי זמין ially, תוך הצגה ברמזי מידע והדרכה ניסוי קשורות במסך המחשב. כאן אנו מתארים את המרכיבים השונים של המערכת שאפשרה לנו להפחית את הזמן הנדרש להשגת את תצורת ההקלטה ולהגדיל באופן משמעותי את הסיכוי להקלטה בהצלחה מספר גדול של תאי עצב בו זמנית.

בעקבות השיטות שתוארו לעיל הצלחנו בביצוע הקלטת כל התא של עד שנים עשר נוירונים בו זמנית, כמעט ואכפיל את המספר הגדול ביותר של תאי עצב בו זמנית ובכך מרחיק הידק התיקון. דוגמאות לרשתות של קשרים סינפטיים ישירים בין פירמידת נוירונים נרשמו בשכבת V של הקליפה החושית של חולדות מו...

Watch this Scientific Journal Video about A Computer-assisted Multi-electrode Patch-clamp System at JoVE.com

A Computer-assisted Multi-electrode Patch-clamp System

הקלטות תיקון מהדק Multi-אלקטרודה מהוות משימה מורכבת. כאן אנו מראים כיצד, על ידי אוטומציה של רבים מהשלבים הניסיוניים, זה אפשרי כדי להאיץ את התהליך מוביל לשיפור איכותי בביצועים ומספר ההקלטות.

מערכת תיקון מהדק Multi-אלקטרודה בסיוע מחשב

The patch-clamp technique is today the most  well-established method for recording electrical activity from individual  neurons or their subcellular ...

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Research

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Neuroscience

13.7K Views.  Ecole Polytechnique Federale de Lausanne.  הקלטות תיקון מהדק Multi-אלקטרודה מהוות משימה מורכבת. כאן אנו מראים כיצד, על ידי אוטומציה של רבים מהשלבים הניסיוניים, זה אפשרי כדי להאיץ את התהליך מוביל לשיפור איכותי בביצועים ומספר ההקלטות. 

Video: A Computer-assisted Multi-electrode Patch-clamp System

Multi-electrode Array Recordings of Human Epileptic Postoperative Cortical Tissue

Epilepsy, affecting about 1% of the population, comprises a group of neurological disorders characterized by the periodic occurrence of seizures, which disrupt normal brain function. Despite treatment with currently available antiepileptic drugs targeting neuronal functions, one third of patients with epilepsy are pharmacoresistant. In this condition, surgical resection of the brain area generating seizures remains the only alternative treatment. Studying human epileptic tissues has contributed to understand new epileptogenic mechanisms during the last 10 years. Indeed, these tissues generate spontaneous interictal epileptic discharges as well as pharmacologically-induced ictal events which can be recorded with classical electrophysiology techniques. Remarkably, multi-electrode arrays (MEAs), which are microfabricated devices embedding an array of spatially arranged microelectrodes, provide the unique opportunity to simultaneously stimulate and record field potentials, as well as action potentials of multiple neurons from different areas of the tissue. Thus MEAs recordings offer an excellent approach to study the spatio-temporal patterns of spontaneous interictal and evoked seizure-like events and the mechanisms underlying seizure onset and propagation. Here we describe how to prepare human cortical slices from surgically resected tissue and to record with MEAs interictal and ictal-like events ex vivo.

We here describe how to perform multi-electrode array recordings of human epileptic cortical tissue. Epileptic tissue resection, slice preparation and multi-electrode array recordings of interictal and ictal events are demonstrated in detail.

הקלטות מערך רב-אלקטרודה של רקמה האנושית אפילפטיים לאחר הניתוח בקליפת המוח

Epilepsy, affecting about 1% of the population, comprises a group of neurological disorders characterized by the periodic occurrence of seizures, which ...

Medicine

Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes

The protocol presented here is designed to study the activation of the large conductance, voltage- and Ca2+-activated K+ (BK) channels. The protocol may also be used to study the structure-function relationship for other ion channels and neurotransmitter receptors1. BK channels are widely expressed in different tissues and have been implicated in many physiological functions, including regulation of smooth muscle contraction, frequency tuning of inner hair cells and regulation of neurotransmitter release2-6. BK channels are activated by membrane depolarization and by intracellular Ca2+ and Mg2+ 6-9. Therefore, the protocol is designed to control both the membrane voltage and the intracellular solution. In this protocol, messenger RNA of BK channels is injected into Xenopus laevis oocytes (stage V-VI) followed by 2-5 days of incubation at 18&deg;C10-13. Membrane patches that contain single or multiple BK channels are excised with the inside-out configuration using patch clamp techniques10-13. The intracellular side of the patch is perfused with desired solutions during recording so that the channel activation under different conditions can be examined. To summarize, the mRNA of BK channels is injected into Xenopus laevis oocytes to express channel proteins on the oocyte membrane; patch clamp techniques are used to record currents flowing through the channels under controlled voltage and intracellular solutions.

Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes

Ionic current of BK channels is recorded using patch clamp techniques. BK channels are expressed in Xenopus oocytes by injecting messenger RNA. The intracellular solution during patch clamp recordings is controlled by a perfusion system.

הצמד תיקון וטכניקות זלוף עבור לימוד תעלות יונים מתבטא Xenopus ביציות

The protocol presented here is designed to study the activation of the large conductance, voltage- and Ca2+-activated K+ (BK) channels. The protocol may ...

Biology

Application of Automated Image-guided Patch Clamp for the Study of Neurons in Brain Slices

Whole-cell patch clamp is the gold-standard method to measure the electrical properties of single cells. However, the in vitro patch clamp remains a challenging and low-throughput technique due to its complexity and high reliance on user operation and control. This manuscript demonstrates an image-guided automatic patch clamp system for in vitro whole-cell patch clamp experiments in acute brain slices. Our system implements a computer vision-based algorithm to detect fluorescently labeled cells and to target them for fully automatic patching using a micromanipulator and internal pipette pressure control. The entire process is highly automated, with minimal requirements for human intervention. Real-time experimental information, including electrical resistance and internal pipette pressure, are documented electronically for future analysis and for optimization to different cell types. Although our system is described in the context of acute brain slice recordings, it can also be applied to the automated image-guided patch clamp of dissociated neurons, organotypic slice cultures, and other non-neuronal cell types.

This protocol describes how to conduct automatic image-guided patch-clamp experiments using a system recently developed for standard in vitro electrophysiology equipment.

יישום אוטומטי של תמונה מודרך תיקון מהדק לחקר נוירונים של פרוסות המוח

Whole-cell patch clamp is the gold-standard method to measure the electrical properties of single cells. However, the in vitro patch clamp remains a ...

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic tectum, is a popular model to study how neural circuits self-assemble. The ability to carry out whole cell patch clamp recordings from tectal neurons and to record RGC-evoked responses, either in vivo or using a whole brain preparation, has generated a large body of high-resolution data about the mechanisms underlying normal, and abnormal, circuit formation and function. Here we describe how to perform the in vivo preparation, the original whole brain preparation, and a more recently developed horizontal brain slice preparation for obtaining whole cell patch clamp recordings from tectal neurons. Each preparation has unique experimental advantages. The in vivo preparation enables the recording of the direct response of tectal neurons to visual stimuli projected onto the eye. The whole brain preparation allows for the RGC axons to be activated in a highly controlled manner, and the horizontal brain slice preparation allows recording from across all layers of the tectum.

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

In this paper, we discuss three brain preparations used for whole cell patch clamp recording to study the retinotectal circuit of Xenopus laevis tadpoles. Each preparation, with its own specific advantages, contributes to the experimental tractability of the Xenopus tadpole as a model to study neural circuit function.

ההכנות ופרוטוקולים עבור כל תא, תיקון קלאמפ הקלטה של נוירונים Tectal צפרדע רפואית זריזה

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic ...

Use of In Vivo Single-fiber Recording and Intact Dorsal Root Ganglion with Attached Sciatic Nerve to Examine the Mechanism of Conduction Failure

Single-fiber recording has been a classical and effective electrophysiological technique over the last few decades because of its specific application for nerve fibers in the central and peripheral nervous systems. This method is particularly applicable to dorsal root ganglia (DRG), which are primary sensory neurons that exhibit a pseudo-unipolar structure of nervous processes. The patterns and features of the action potentials passed along axons are recordable in these neurons. The present study uses in vivo single-fiber recordings to observe the conduction failure of sciatic nerves in complete Freund&#8217;s adjuvant (CFA)-treated rats. As the underlying mechanism cannot be studied using in vivo single-fiber recordings, patch-clamp-recordings of DRG neurons are performed on preparations of intact DRG with the attached sciatic nerve. These recordings reveal a positive correlation between conduction failure and the rising slope of the after-hyperpolarization potential (AHP) of DRG neurons in CFA-treated animals. The protocol for in vivo single fiber-recordings allows the classification of nerve fibers via the measurement of conduction velocity and monitoring of abnormal conditions in nerve fibers in certain diseases. Intact DRG with attached peripheral nerve allows observation of the activity of DRG neurons in most physiological conditions. Conclusively, single-fiber recording combined with electrophysiological recording of intact DRGs is an effective method to examine the role of conduction failure during the analgesic process.

Single-fiber recording is an effective electrophysiological technique that is applicable to the central and peripheral nervous systems. Along with the preparation of intact DRG with the attached sciatic nerve, the mechanism of conduction failure is examined. Both protocols improve the understanding of the peripheral nervous system&#39;s relationship with pain.

השימוש ב Vivo הקלטה חד-סיבים ושורש השורש שלמות עם העצב מוצמד לבחון את המנגנון של כשל הולכה

Single-fiber recording has been a classical and effective electrophysiological technique over the last few decades because of its specific application for ...

One-channel Cell-attached Patch-clamp Recording

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate depends on properties intrinsic to each channel protein as well as the mechanism by which it is generated and controlled and represents an important area of current research. Information about the operational dynamics of ion channel proteins can be obtained by observing long stretches of current produced by a single molecule. Described here is a protocol for obtaining one-channel cell-attached patch-clamp current recordings for a ligand gated ion channel, the NMDA receptor, expressed heterologously in HEK293 cells or natively in cortical neurons. Also provided are instructions on how to adapt the method to other ion channels of interest by presenting the example of the mechano-sensitive channel PIEZO1. This method can provide data regarding the channel&#8217;s conductance properties and the temporal sequence of open-closed conformations that make up the channel&#8217;s activation mechanism, thus helping to understand their functions in health and disease.

Described here is a procedure for obtaining long stretches of current recording from one ion channel with the cell-attached patch-clamp technique. This method allows for observing, in real time, the pattern of open-close channel conformations that underlie the biological signal. These data inform about channel properties in undisturbed biological membranes.

הקלטת תיקון מהדק מצורף תא אחד ערוצים

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate ...

Patch Clamp Electrophysiology

Neuron cell membranes are populated with ion channels that control the movement of charge into and out of the cell, thereby regulating neuron firing. One extremely useful technique for investigating the biophysical properties of these channels is called patch clamp recording. In this method, neuroscientists place a polished glass micropipette against a cell and apply suction to form a high resistance seal. This process isolates a small &#8220;patch&#8221; of membrane that contains one or more ion channels. Using an electrode within the micropipette, researchers can &#8220;clamp&#8221; or control the electrical properties of the membrane, which is important for analysis of channel activity. The electrode also allows for changes in the voltage across the membrane, or the flow of ions through the membrane, to be recorded. This video begins with an overview of the principles behind patch clamp electrophysiology, an introduction to the necessary equipment, and descriptions of the various patch configurations, including whole cell, cell-attached, perforated, inside-out, and outside-out patches. Next, the key steps of a typical whole-cell patch clamp experiment are outlined, in which a current-voltage (IV) curve is generated. Finally, applications of patch clamp recording are provided to demonstrate how the biophysical properties of ion channels, cell excitability, and neuroactive compounds are evaluated in neurophysiology labs today.

Patch Clamp Electrophysiology: Principle &amp; Applications

אלקטרופיזיולוגיה של מהדק תיקון

Neuron cell membranes are populated with ion channels that control the movement of charge into and out of the cell, thereby regulating neuron firing. One ...

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Advanced Biology

Establishing a Whole-Cell Configuration through the Patch Clamp Method

Source: Segev, A. et al., <a href="https://app.jove.com/v/54024">Whole-cell Patch-clamp Recordings in Brain Slices.</a>&#160;J. Vis. Exp.&#160;(2016)
This video demonstrates the whole-cell patch clamp technique in a mouse brain slice. This technique establishes a stable connection between micropipette and neuron membrane, which is further disrupted to establish a whole-cell configuration to study the neuron&#39;s health and physiological responses.

יצירת תצורת תא שלם באמצעות שיטת Patch Clamp

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
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JoVE Encyclopedia of Experiments

Neurophysiology

Electrophysiology Techniques

Amphotericin-B Mediated Perforated Patch-Clamp Analysis: An Electrophysiological Technique to Study Ionic Currents in Urinary Bladder Cells

Source: Malysz, J. et al. <a href="https://www.jove.com/video/59884" target="_blank">Preparation and Utilization of Freshly Isolated Human Detrusor Smooth Muscle Cells for Characterization of 9-Phenanthrol-Sensitive Cation Currents.</a> J. Vis. Exp. (2020)
This video demonstrates the characterization of cationic currents from transient receptor potential melastatin type 4 (TRPM4) ion channels in detrusor smooth muscle cells using amphotericin-B mediated perforated patch-clamp technique. The technique helps evaluate the biophysical and pharmacological properties of ion channels.

Amphotericin-B מתווך תיקון מהדק תיקון: טכניקה אלקטרופיזיולוגית לחקר זרמים יוניים בתאי שלפוחית השתן

Source: Malysz, J. et al. Preparation and Utilization of Freshly Isolated Human Detrusor Smooth Muscle Cells for Characterization of ...

Cancer Research

Urinary Tract Cancer

In vitro studies

Whole-Cell Patch Clamp Electrophysiology: A Method to Study Electrical Properties of Neurons

Source: Roy, B., et al. <a href="https://www.jove.com/v/50551/" target="_blank">Patch Clamp Recordings from Embryonic Zebrafish Mauthner Cells</a>. J. Vis. Exp. (2013).
This protocol describes a technique called whole-cell patch clamp electrophysiology, which is a method to study electrical properties of Mauthner neurons and other reticulospinal cells in zebrafish embryos.

אלקטרופיזיולוגיה של מהדק טלאי תאי שלם: שיטה לחקר תכונות חשמליות של נוירונים

Source: Roy, B., et al. Patch Clamp Recordings from Embryonic Zebrafish Mauthner Cells. J. Vis. Exp. (2013).
This protocol describes a technique called ...