Name: single-cell suction recordings from mouse cone photoreceptors
Uploaded: 2010-01-05T20:00:00
Description: Watch this Scientific Journal Video about Single-cell Suction Recordings from Mouse Cone Photoreceptors at JoVE.com

从研究职业发展奖从研究的支撑，防止失明，美国国立卫生研究院授予安永019312，和不受限制的赠款，防止盲目性和安永02687（华盛顿大学眼科及视觉科学系）。

制作电极<ol><li>请使用微量拉马的记录电极和波兰与加热丝在显微镜下的电极。鼠标锥单细胞吸记录，电极尖端内径约6-7微米。 </li><li>准备参比电极。称取蒸馏水1％琼脂。在热水浴融化的琼脂溶液。用1％琼脂使用注射器填充玻璃吸液管（L = 100毫米，外径/ ID = 1/0.75毫米）。巩固在室温下10分钟的琼脂。 </li><li>切成半使用钻石刀的移液器。 </li><li>对照品溶液的参比电极浸泡至少24小时在4 ° C。 </l...

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	<li>Yau, K. W., Lamb, T. D., Baylor, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Light-induced+fluctuations+in+membrane+current+of+single+toad+rod+outer+segments.">Light-induced fluctuations in membrane current of single toad rod outer segments.</a> Nature. 269, 78-80 (1977).</li><li>Nikonov, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoreceptors+of+Nrl+-/-+mice+coexpress+functional+S-+and+M-cone+opsins+having+distinct+inactivation+mechanisms.">Photoreceptors of Nrl -/- mice coexpress functional S- and M-cone opsins having distinct inactivation mechanisms.</a> J Gen Physiol. 125, 287-304 (2005).</li><li>Nikonov, S. S., Kholodenko, R., Lem, J., Pugh, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiological+features+of+the+S-+and+M-cone+photoreceptors+of+wild-type+mice+from+single-cell+recordings.">Physiological features of the S- and M-cone photoreceptors of wild-type mice from single-cell recordings.</a> J Gen Physiol. 127, 359-374 (2006).</li><li>Applebury, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+murine+cone+photoreceptor:+a+single+cone+type+expresses+both+S+and+M+opsins+with+retinal+spatial+patterning.">The murine cone photoreceptor: a single cone type expresses both S and M opsins with retinal spatial patterning.</a> Neuron. 27, 513-523 (2000).</li><li>Cornwall, M. C., Fein, A., MacNichol, E. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+mechanisms+that+underlie+bleaching+and+background+adaptation.">Cellular mechanisms that underlie bleaching and background adaptation.</a> J Gen Physiol. 96, 345-372 (1990).</li><li>Calvert, P. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phototransduction+in+transgenic+mice+after+targeted+deletion+of+the+rod+transducin+alpha+-subunit.">Phototransduction in transgenic mice after targeted deletion of the rod transducin alpha -subunit.</a> Proc Natl Acad Sci U S A. 97, 13913-13918 (2000).</li></ol>

发展三十年前从感光细胞的单细胞吸录音。它使我们能够记录光刺激引起的不穿透细胞膜的跨膜电流变化。由于高的细胞与细胞间的粘附，它是很难隔离鼠标像两栖类动物的视网膜视网膜健康的单杆和锥，它是很难找到的比例很低（3％）和小尺寸，由于个人锥。段内（IS - 在）记录方法，记录从目前的几个感光器，其中可能包括一个圆锥体的内在环节，克服了这一困难。在野生型小鼠，反应应该?...

single-cell suction recordings from mouse cone photoreceptors

在视网膜上的杆和锥光感受器负责光检测。在黑暗中，环核苷酸门控技术（CNG）外段的渠道是开放的，允许离子流稳步向内穿过细胞膜，去极化的细胞。光线照射触发关闭的CNG通道，阻止外来阳离子电流的流动，从而导致细胞超极化。基于光感受器的极性，吸记录方法是发达国家在20世纪70年代，不像传统的膜片钳技术，不需要穿透质膜<sup&gt; 1</sup&gt;。绘制成一个紧密拟合的玻璃吸管充满了细胞外液的外节，可以录制测试闪光曝光后单个细胞的电流变化。但是，这种行之有效的“外段（操作系统）”吸气录制不适合鼠标锥录音，因为在小鼠视网膜（3％）和确定锥体困难的视锥细胞的比例很低外节。近日，在段内（中）录制的配置吸引到录音吸液管内段/核区的感光<sup&gt; 2,3</sup&gt;。在这段视频中，我们将展示如何从个别鼠标使用单细胞吸电极锥photoresponses记录。

Watch this Scientific Journal Video about Single-cell Suction Recordings from Mouse Cone Photoreceptors at JoVE.com

Single-cell Suction Recordings from Mouse Cone Photoreceptors

我们将展示如何从单一的鼠标采用吸电极锥记录闪光的反应。

单从小鼠锥感光细胞吸录音

Rod and cone photoreceptors in the retina are responsible for light detection. In darkness, cyclic nucleotide-gated (CNG) channels in the outer segment ...

Research

JoVE Journal

Biology

Methods for Patch Clamp Capacitance Recordings from the Calyx

膜片钳从花萼电容录音方法

We demonstrate the basic techniques for presynaptic patch clamp recording at the calyx of Held, a mammalian central nervous system nerve terminal. ...

科研

生物学

Whole Cell Recordings from Brain of Adult Drosophila

从整个成年果蝇大脑细胞记录

In this video, we demonstrate the procedure for isolating whole brains from adult Drosophila in preparation for recording from single neurons. We begin by ...

Preparation of Single-Cell Suspensions from Mouse Spleen with the gentleMACS Dissociator

gentleMACS Dissociator从小鼠脾脏单细胞悬浮液的制备

Single-cell suspensions are a prerequisite for experiments in cell separation, cell analysis and cell culture.  To avoid tedious and often painful manual ...

Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator

标准化的制备小鼠肺组织单细胞悬浮液使用gentleMACS Dissociator

The preparation of single-cell suspensions from tissues is an important prerequisite for many experiments in cellular research. The process of ...

Generation of Single-Cell Suspensions from Mouse Neural Tissue

从小鼠神经组织产生单细胞悬浮液

Within the nervous system, hundreds of neuronal and glial cell types have been described. Each specific cell type in the brain or spinal cord has a ...

Single Cell Transcriptional Profiling of Adult Mouse Cardiomyocytes

成年小鼠心肌细胞的单细胞转录谱

While numerous studies have examined gene expression changes from homogenates of heart tissue, this prevents studying the inherent stochastic variation ...

Generation of Mice Derived from Induced Pluripotent Stem Cells

诱导多能干细胞来源于小鼠的生成

The production of induced pluripotent stem cells (iPSCs) from somatic cells provides a means to create valuable tools for basic research and may also ...

Isolation of Endothelial Cells from the Lumen of Mouse Carotid Arteries for Single-Cell Multi-Omics Experiments

Atherosclerosis is an inflammatory disease of the arterial regions exposed to disturbed blood flow (d-flow). D-flow regulates the expression of genes in ...

Isolation of Nuclei from Flash-Frozen Liver Tissue for Single-Cell Multiomics

从快速冷冻肝组织中分离细胞核用于单细胞多组学

The liver is a complex and heterogenous tissue responsible for carrying out many critical physiological functions, such as the maintenance of energy ...

Single-Cell Suspension Preparation from Nile Tilapia Intestine for Single-Cell Sequencing

尼罗罗非鱼肠单细胞悬液制备用于单细胞测序

Nile tilapia is one of the most commonly cultured freshwater fish species worldwide and is a widely used research model for aquaculture fish studies. The ...