Name: calcium imaging of cortical neurons using fura-2 am
Uploaded: 2009-01-19T00:00:00
Description: Watch this Scientific Journal Video about Calcium Imaging of Cortical Neurons using Fura-2 AM at JoVE.com

<p class="jove_title">细胞培养</p><p class="jove_content">细胞可以种植使用成熟的技术，但必须在1＃玻璃涂上与蜂窝胶粘剂（如聚赖氨酸，polyornithine或层粘连蛋白），以防止细胞分离或在移动成像实验盖玻片镀。 </p><p class="jove_title">解决方案</p><p class="jove_content">可以使用各种生理的解决方案，包括细胞培养基钙成像实验。然而，重要的是，以确保该解决方案是无酚红，这大大增加了荧光背景。我们使用Tyrodes解决方案，这?...

<ol>
	<li>Grynkiewicz, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+generation+of+Ca2++indicators+with+greatly+improved+fluorescence+properties.">A new generation of Ca2+ indicators with greatly improved fluorescence properties.</a> <em style='font-style: italic'>J. Biol. Chem</em>. <b>260</b>, 3440-3450 (1985).</li><li>Lewis, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitogen-induced+oscillations+of+cytosolic+Ca2++and+transmembrane+Ca2++current+in+human+leukemic+T+cells.">Mitogen-induced oscillations of cytosolic Ca2+ and transmembrane Ca2+ current in human leukemic T cells.</a> <em style='font-style: italic'>Cell Regul</em>. <b>1</b>, 99-112 (1989).</li><li>Dolmetsch, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Signaling+between+intracellular+Ca2++stores+and+depletion-activated+Ca2++channels+generates+[Ca2+]i+oscillations+in+T+lymphocytes.">Signaling between intracellular Ca2+ stores and depletion-activated Ca2+ channels generates [Ca2+]i oscillations in T lymphocytes.</a> <em style='font-style: italic'>J Gen Physiol</em>. <b>103</b>, 365-388 (1994).</li></ol>

<p>在此演示中，我们已经经历了使用FURA - 2上午进行钙成像的所有步骤。我们作为一个模型的皮层神经元，但可用于实时测量多种细胞的细胞内钙离子钙成像。当执行此过程，重要的是用盖玻片代替塑料，因为塑料是经常在荧光紫外线的波长，这使得成像困难。此外，重要的是预涂层与polyornithine和层粘连蛋白的盖玻片，以防止从分离的细胞。最后，重要的是要记住以避免产生气泡的解决方案，通过成像室时灌流。 </p>

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<td>Warner Instruments</td>
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calcium imaging of cortical neurons using fura-2 am

<p class="jove_content">钙成像是一种常见的的技术，用于测量培养细胞中的钙信号。钙成像技术的优势，这是BAPTA为基础的有机分子，Ca2 +离子结合，改变其光谱特性的钙染料。钙染料分为两大类，如FURA - 2和Indo - 1和单波长染料如荧光4的比例度量的染料。比率度量染料改变他们的激发或发射光谱响应钙，使细胞内钙离子浓度在不同的波长的荧光发射或激发的比例确定。使用单波长探测器的比例度量的染料的主要优势是，信噪比是独立的染料浓度，光照强度，和光路长度，使细胞内钙离子的浓度，以确定这些文物的独立。最常见的钙指标之一是FURA - 2，它已经在505 nm处的发射峰从340 nm处的激发峰，380 nm的钙结合。在这里，我们描述了使用FURA - 2，在神经元和其他可兴奋细胞来衡量细胞内钙升高。</p>

Watch this Scientific Journal Video about Calcium Imaging of Cortical Neurons using Fura-2 AM at JoVE.com

Calcium Imaging of Cortical Neurons using Fura-2 AM

<p class="jove_content">钙信号，发挥了关键作用，在许多细胞过程，包括基因表达，存活和分化。在这里，我们将演示如何执行使用FURA - 2的钙成像。钙成像是一种宝贵的的工具来研究细胞内钙离子的实时调控及其信号传导的调控。</p>

使用FURA - 2上午皮层神经元的钙成像

Calcium imaging is a common technique that is useful for measuring calcium signals in cultured cells. Calcium imaging techniques take advantage of calcium ...

Research

JoVE Journal

Biology

Preparing E18 Cortical Rat Neurons for Compartmentalization in a Microfluidic Device

准备在微流体装置E18条块皮质大鼠神经元

In this video, we demonstrate the preparation of E18 cortical rat neurons. E18 cortical rat neurons are obtained from E18 fetal rat cortex previously ...

科研

生物学

In Vivo 2-Photon Calcium Imaging in Layer 2/3 of Mice

在体内2光子层钙成像对小鼠的2 / 3

To understand network dynamics of microcircuits in the neocortex, it is essential to simultaneously record the activity of a large number of neurons .  ...

Live Imaging of Dense-core Vesicles in Primary Cultured Hippocampal Neurons

在原代培养海马神经元的密集核心囊泡的实时成像

Observing and characterizing dynamic cellular processes can yield important information about cellular activity that cannot be gained from static images.  ...

<em>In vivo</em> Imaging of Deep Cortical Layers using a Microprism

In vivo Imaging of Deep Cortical Layers using a Microprism

<em>在</em>使用微深皮质层<em>的活体</em>成像

We present a protocol for in vivo imaging of cortical tissue using a deep-brain imaging probe in the shape of a microprism.  Microprisms are 1-mm in size ...

Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor

使用一种基因编码的钙传感器来监测细胞凋亡过程中线粒体钙水平的动态变化

Dynamic changes in intracellular calcium concentration in response to various stimuli regulates many cellular processes such as proliferation, ...

Fluorescence-based Measurement of Store-operated Calcium Entry in Live Cells: from Cultured Cancer Cell to Skeletal Muscle Fiber

基于荧光测定活细胞中的钙进入商店经营：从培养的肿瘤细胞，骨骼肌纤维

Store operated Ca2+ entry (SOCE), earlier termed capacitative Ca2+ entry, is a tightly regulated mechanism for influx of extracellular Ca2+ into cells to ...

Direct Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading (TED)

Direct Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading TED

直接成像诱导ER钙有针对性的酯酶染料装载（TED）

Visualization of calcium dynamics is important to understand the role of calcium in cell physiology. To examine calcium dynamics, synthetic fluorescent ...

Measurement of Calcium Fluctuations Within the Sarcoplasmic Reticulum of Cultured Smooth Muscle Cells Using FRET-based Confocal Imaging

基于FRET钙内波动培养平滑肌细胞使用的肌质网的测量共焦成像

Maintenance of steady-state calcium (Ca2+) levels in the sarcoplasmic reticulum (SR) of vascular smooth muscle cells (VSMCs) is vital to their overall ...

Concurrent EEG and Functional MRI Recording and Integration Analysis for Dynamic Cortical Activity Imaging

动态皮层活动影像的并发脑电图与功能 MRI 记录及积分分析

Electroencephalography (EEG) and functional magnetic resonance imaging (fMRI) are two of the fundamental noninvasive methods for identifying brain ...

Analysis of Beta-cell Function Using Single-cell Resolution Calcium Imaging in Zebrafish Islets

斑马鱼胰岛单细胞分辨率钙成像对β细胞功能的分析

Pancreatic beta-cells respond to increasing blood glucose concentrations by secreting the hormone insulin. The dysfunction of beta-cells leads to ...