Name: long-term culture and monitoring of isolated caenorhabditis elegans on solid media in multi-well devices
Uploaded: 2022-12-09T14:10:00
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这项工作得到了NIH R35GM133588对GLS，美国国家医学院催化剂奖对GLS，亚利桑那州技术和研究倡议基金的支持，由亚利桑那州董事会和埃里森医学基金会管理。

1. 储备溶液和培养基的制备
注意：在开始制备多孔设备之前，请准备以下储备溶液和培养基。
<ol><li>线虫生长培养基 （NGM） 和低熔点 NGM （lmNGM） 的储备溶液：<ol><li>准备 1 M K 2 HPO4：将 174.18 gK 2HPO4 加入1 L 瓶中，并用无菌去离子水填充至 1 L。高压灭菌器（121°C，15psig）30分钟，并在室温（RT）下储存。</li>
		<li>准备 1 M KPi，pH 6.0...

<ol>
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WorMotel系统是一个强大的工具，可以随着时间的推移收集数百个孤立的 秀丽隐杆线虫 的个性化数据。继早期使用多孔装置应用于发育静止、运动行为和衰老的研究之后，这项工作的目标是优化多孔装置的制备，以便以更高的通量方式长期监测活动、健康和寿命。这项工作提供了制备多孔设备的详细协议，该协议优化了原始协议13的许多步骤，突出了可能存在技术困难的关?...

秀丽隐杆线虫 通常用于衰老研究，因为它们的生成时间短（约3天），寿命短（约3周），易于在实验室中培养，与哺乳动物的分子过程和途径的高度进化保守，以及遗传操作技术的广泛可用性。在衰老研究的背景下， 秀丽隐杆线虫 允许快速生成长寿数据和老年人口，以分析活体动物的晚年表型。进行蠕虫老化研究的典型方法包括在6cm培养皿1中手动测量固体琼脂线虫生长培养基（NGM）上以20至70只动物为一组的蠕虫群的寿命。使用年龄同步的种群可以测量整个种群中个体动物的寿命或横截面表型，但这种方法排除了随着时间的推移监测个体动物的特征。这种方法也是劳动密集型的，因此限制了可以测试的人口规模。
允许在秀丽隐杆线虫的整个生命周期中纵向监测的培养方法数量有限，每种方法都有一系列独特的优点和缺点。微流体设备，包括WormFarm2，NemaLife3和“行为”芯片4，以及5，6，7，允许随着时间的推移监测个体动物。使用多孔板在液体培养物中培养蠕虫同样允许随着时间的推移监测单个动物或少量秀丽隐杆线虫种群8，9。液体环境代表了与培养皿中固体培养基上的常见培养环境不同的环境背景，这可以改变与衰老相关的动物生理学方面，包括脂肪含量和应激反应基因的表达10，11。将这些研究与收集的大多数关于老年秀丽隐杆线虫的数据直接进行比较的能力受到潜在重要环境变量差异的限制。蠕虫畜栏12是一种在更接近于复制典型固体培养基培养物的环境中容纳个体动物的方法。蠕虫畜栏在使用水凝胶的显微镜载玻片上为每只动物包含一个密封室，可以对孤立的动物进行纵向监测。该方法使用标准明场成像来记录形态数据，例如体型和活动。然而，动物作为胚胎被放置在水凝胶环境中，在那里它们在整个生命周期中不受干扰。这需要使用有条件的无菌突变体或转基因遗传背景，这既限制了遗传筛选的能力，因为每个新的突变或转基因都需要杂交到具有条件不育的背景中，以及药物筛选的能力，因为治疗只能作为胚胎应用于动物一次。
Fang-Yen实验室开发的另一种方法允许在称为WorMotel13，14的微加工聚二甲基硅氧烷（PDMS）装置的单个孔中的固体培养基上培养蠕虫。每个装置都放置在单孔托盘中（即，尺寸与96孔板相同），并具有240个孔，由充满厌恶溶液的护城河隔开，以防止蠕虫在孔之间移动。每口井在其生命周期内可以容纳一个蠕虫。该设备被吸水聚丙烯酰胺凝胶颗粒（称为“水晶体”）包围，托盘用石蜡膜实验室薄膜密封，以保持湿度并最大限度地减少介质的干燥。该系统允许收集个体动物的健康寿命和寿命数据，而固体培养基的使用可以更好地概括绝大多数已发表的 秀丽隐杆线虫 寿命研究中动物所经历的环境，从而允许更直接的比较。最近，已经使用最初用于微细胞毒性测定15 的聚苯乙烯微托盘代替PDMS装置16开发了类似的技术。微托盘方法允许收集在固体培养基上培养的蠕虫的个性化数据，并提高了在通常会导致逃跑的条件下（例如，压力源或饮食限制）容纳蠕虫的能力，权衡是每个微托盘只能包含96只动物16，而这里使用的多孔装置最多可以容纳240只动物。
这里介绍的是用于制备多孔设备的详细方案，该方案针对板对板的一致性和并行制备多个设备进行了优化。该协议改编自方颜实验室13的原始协议。具体来说，有关于最小化污染、优化固体培养基和细菌食物源的一致干燥以及输送RNAi和药物的技术的描述。该系统可用于跟踪个人健康寿命、寿命和其他表型，例如体型和形状。这些多孔设备与现有的高通量系统兼容，可以测量寿命，这可以消除传统寿命实验中涉及的大部分体力劳动，并为大规模对单个 秀丽隐杆线虫 进行自动化、直接的寿命测量和健康跟踪提供机会。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>2.5 lb weight</td>
			<td>CAP Barbell</td>
			<td>RP-002.5</td>
			<td></td>
		</tr>
		<tr>
			<td>Acrylic sheets (6 in x 4 in x 3/8 in)</td>
			<td>Falken Design</td>
			<td>ACRYLIC-CL-3-8/1224</td>
			<td>Large sheet cut to smaller sizes&#160;</td>
		</tr>
		<tr>
			<td>Ampicillin sodium salt</td>
			<td>Sigma-Aldrich</td>
			<td>A9518</td>
			<td></td>
		</tr>
		<tr>
			<td>Autoclavable squeeze bottle</td>
			<td>Nalgene</td>
			<td>2405-0500</td>
			<td></td>
		</tr>
		<tr>
			<td>Bacto agar</td>
			<td>BD Difco</td>
			<td>214030</td>
			<td></td>
		</tr>
		<tr>
			<td>Bacto peptone</td>
			<td>Thermo Scientific</td>
			<td>211677</td>
			<td></td>
		</tr>
		<tr>
			<td>Basin, 25 mL</td>
			<td>VWR</td>
			<td>89094-664</td>
			<td>Disposable pipette basin&#160;</td>
		</tr>
		<tr>
			<td>Cabinet style vacuum desiccator&#160;</td>
			<td>SP Bel-Art</td>
			<td>F42400-4001</td>
			<td>Do not need to use dessicant, only using as a vacuum chamber.&#160;</td>
		</tr>
		<tr>
			<td>CaCl2</td>
			<td>Acros Organics</td>
			<td>349615000</td>
			<td></td>
		</tr>
		<tr>
			<td>Caenorhabditis elegans N2</td>
			<td>Caenorhabditis Genetics Center (CGC)</td>
			<td>N2</td>
			<td>Wildtype strain</td>
		</tr>
		<tr>
			<td>Carbenicillin&#160;</td>
			<td>GoldBio</td>
			<td>C-103-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Beckman</td>
			<td>360902</td>
			<td></td>
		</tr>
		<tr>
			<td>Cholesterol</td>
			<td>ICN Biomedicals Inc</td>
			<td>101380</td>
			<td></td>
		</tr>
		<tr>
			<td>Compressed oxygen tank</td>
			<td>Airgas</td>
			<td>UN1072</td>
			<td></td>
		</tr>
		<tr>
			<td>CuSO4</td>
			<td>Fisher Chemical</td>
			<td>C493-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Dry bead bath incubator</td>
			<td>Fisher Scientific</td>
			<td>11-718-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Escherichia coli OP50&#160;</td>
			<td>Caenorhabditis Genetics Center (CGC)</td>
			<td>OP50</td>
			<td>Standard labratory food for C. elegans</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Millipore</td>
			<td>ex0276-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Floxuridine</td>
			<td>Research Products International</td>
			<td>F10705-1.0</td>
			<td></td>
		</tr>
		<tr>
			<td>Hybridization oven</td>
			<td>Techne</td>
			<td>731-0177</td>
			<td>Used to cure PDMS mixture, any similar oven will suffice</td>
		</tr>
		<tr>
			<td>Incubators</td>
			<td>Shel Lab</td>
			<td>2020</td>
			<td>20 &#176;C incubator for maintaining worm strains and 37 &#176;C incubator to grow bacteria&#160;</td>
		</tr>
		<tr>
			<td>Isopropyl &#223;-D-1-thiogalactopyranoside (IPTG)</td>
			<td>GoldBio</td>
			<td>I2481C100</td>
			<td></td>
		</tr>
		<tr>
			<td>K2HPO4</td>
			<td>Fisher Chemical</td>
			<td>P288-500</td>
			<td></td>
		</tr>
		<tr>
			<td>KH2PO4</td>
			<td>Fisher Chemical</td>
			<td>P286-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Kimwipes</td>
			<td>KimTech</td>
			<td>34155</td>
			<td>Task wipes</td>
		</tr>
		<tr>
			<td>LB Broth, Lennox</td>
			<td>BD Difco</td>
			<td>240230</td>
			<td></td>
		</tr>
		<tr>
			<td>Low melt agarose</td>
			<td>Research Products International</td>
			<td>A20070-250.0</td>
			<td></td>
		</tr>
		<tr>
			<td>MgSO4</td>
			<td>Fisher Chemical</td>
			<td>M-8900</td>
			<td></td>
		</tr>
		<tr>
			<td>Microwave&#160;</td>
			<td>Sharp</td>
			<td>R-530DK</td>
			<td></td>
		</tr>
		<tr>
			<td>Multichannel repeat pipette, 20&#8211;200 &#181;L LTS EDP3</td>
			<td>Rainin</td>
			<td>17013800</td>
			<td>The exact model used is no longer sold, a similar model&#39;s catalog number has been provided</td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Fisher Bioreagents</td>
			<td>BP358-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Nunc OmniTray</td>
			<td>Thermo Scientific</td>
			<td>264728</td>
			<td>Clear polystyrene trays</td>
		</tr>
		<tr>
			<td>Parafilm M</td>
			<td>Fisher Scientific</td>
			<td>13-374-10</td>
			<td>Double-wide (4 in)</td>
		</tr>
		<tr>
			<td>Petri plate, 100 mM&#160;</td>
			<td>VWR</td>
			<td>25384-342</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri plate, 60 mM&#160;</td>
			<td>Fisher Scientific</td>
			<td>FB0875713A</td>
			<td></td>
		</tr>
		<tr>
			<td>Plasma cleaner</td>
			<td>Plasma Etch, Inc.</td>
			<td>PE-50</td>
			<td></td>
		</tr>
		<tr>
			<td>PLATINUM vacuum pump</td>
			<td>JB Industries</td>
			<td>DV-142N&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>PolyJet 3D printer</td>
			<td>Stratasys&#160;</td>
			<td>Objet500 Connex3</td>
			<td>PolyJet 3D printing services provided by ProtoCAM (Matrial: Vero Rigid; Finish: Matte; Color: Gloss; Resolution: X-axis: 600 dpi, Y-axis: 600 dpi, Z-axis: 1600 dpi)</td>
		</tr>
		<tr>
			<td>Shaking incubator</td>
			<td>Lab-Line</td>
			<td>3526CC</td>
			<td></td>
		</tr>
		<tr>
			<td>smartSpatula</td>
			<td>LevGo, Inc.</td>
			<td>17211</td>
			<td>Disposable spatula</td>
		</tr>
		<tr>
			<td>Superabsorbent polymer (AgSAP Type S)</td>
			<td>M2 Polymer Technologies</td>
			<td>Type S</td>
			<td>Referred to in main text as &#34;water crystals&#34;</td>
		</tr>
		<tr>
			<td>SYLGARD 184 Silicone Elastomer base</td>
			<td>The Dow Chemical Company</td>
			<td>2065622</td>
			<td></td>
		</tr>
		<tr>
			<td>SYLGARD 184 Silicone Elastomer curing agent</td>
			<td>The Dow Chemical Company</td>
			<td>2085925</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe filter (0.22 &#181;m)</td>
			<td>Nest Scientific USA Inc.&#160;</td>
			<td>380111</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe, 10 mL&#160;</td>
			<td>Fisher Scientific</td>
			<td>14955453</td>
			<td></td>
		</tr>
		<tr>
			<td>TWEEN 20</td>
			<td>Thermo Scientific</td>
			<td>J20605-AP</td>
			<td>Detergent</td>
		</tr>
		<tr>
			<td>Vacuum pump oil</td>
			<td>VWR</td>
			<td>54996-082</td>
			<td></td>
		</tr>
		<tr>
			<td>VeroBlackPlus</td>
			<td>Stratasys&#160;</td>
			<td>RGD875</td>
			<td>Rigid 3D printing filament</td>
		</tr>
		<tr>
			<td>Weigh boat</td>
			<td>Thermo Scientific</td>
			<td>WB30304</td>
			<td>Large enough for PDMS mixture volume</td>
		</tr>
	</tbody>
</table>

long-term culture and monitoring of isolated caenorhabditis elegans on solid media in multi-well devices

线虫秀 丽隐杆线虫 是衰老研究中最常用的模型系统之一，因为它的培养技术简单且便宜，繁殖周期快（~3天），寿命短（~3周），以及许多可用的遗传操作和分子分析工具。在 秀丽隐杆线虫中进行衰老研究的最常见方法（包括生存分析）涉及在培养皿中的固体线虫生长培养基（NGM）上一起培养数十至数百只动物的种群。虽然这种方法收集了动物种群的数据，但大多数协议不会随着时间的推移跟踪个体动物。这里介绍的是在称为WorMotels的微加工聚二甲基硅氧烷（PDMS）设备上长期培养个体动物的优化方案。每个装置允许在含有NGM的小孔中培养多达240只动物，每个孔由含硫酸铜的护城河隔离，以防止动物逃跑。本文以原始 WorMotel 描述为基础，提供了用于成型、准备和填充每个设备的详细协议，并描述了常见的技术复杂性和故障排除建议。在该协议中，有用于一致加载小容量NGM的技术，NGM和细菌食品的一致干燥，提供药物干预的选项，重复使用PDMS设备的说明和实际限制，以及即使在低湿度环境中也能最大限度地减少干燥的技巧。该技术允许在类似于培养皿中固体培养基上群体培养的标准技术的环境中纵向监测各种生理参数，包括刺激活动、未刺激活动、体型、运动几何形状、健康跨度和生存。当与自动显微镜和分析软件结合使用时，该方法与高通量数据收集兼容。最后，讨论了该技术的局限性，并将该方法与最近开发的方法进行了比较，该方法使用微托盘在固体培养基上培养分离的线虫。

WorMotel培养系统可用于收集各种数据，包括有关寿命，健康寿命和活动的数据。已发表的研究利用多孔设备来研究寿命和健康寿命13，14，静止和睡眠22，23，24以及行为25。寿命可以手动评分，也可以通过图像集合和下游成像分析进行评分。在前一种方法中，可以在?...

Watch this Scientific Journal Video about Long-Term Culture and Monitoring of Isolated Caenorhabditis elegans on Solid Media in Multi-Well Devices at JoVE.com

Long-Term Culture and Monitoring of Isolated Caenorhabditis elegans on Solid Media in Multi-Well Devices

这里介绍的是一种优化的方案，用于在微加工多孔设备中的固体培养基上培养分离的单个线虫。这种方法允许在个体动物的一生中监测与衰老和健康相关的各种表型，包括活动、体型和形状、运动几何形状和存活率。

多孔设备固体培养基上分离秀 丽隐杆线虫 的长期培养和监测

The nematode Caenorhabditis elegans is among the most common model systems used in aging research owing to its simple and inexpensive culture techniques, ...

当我们研究衰老和海秀丽隐杆线虫时，我们通常会随着时间的推移跟踪一群动物。这让我们瞥见了你正在研究的过程在每个时间点在种群中做了什么，但它并没有告诉你这个过程是如何随着时间的推移在个体动物中变化的。这些单一动物培养环境使我们能够并行跟踪数百只动物的个体动物分子过程和生理过程随时间变化的动态变化。我们发现，有两件主要因素可以真正影响这些多孔培养系统的质量。一个是井干涸或干涸不足，二是污染。我们尝试在整个协议中注意您可以尝试缓解这些问题的特定步骤。今天展示协议的将是艾米丽·加迪亚。她是我实验室的技术人员，为优化这个协议做了很多工作。准备好所有试剂后，使用一次性刮刀在大型一次性船上混合每个模具60克PDMS基料和6克固化剂。将PDMS混合物置于真空室中，在零下0.08兆帕斯卡下放置30分钟以除去气泡。将PDMS混合物倒入并填充到每个3D打印模具中。将填充的模具和任何额外的PDMS混合物放入真空室中，以零下0.08兆帕斯卡的温度放置25分钟。从真空室中取出填充的模具，并用一次性刮刀弹出任何剩余的气泡。从一个边缘开始，在PDMS填充模具的顶部慢慢降低平坦的亚克力板。如果形成气泡，请按照手稿中的说明进行操作。将 2.5 磅重的重量放在丙烯酸上，然后将加重的模具放入烤箱中。让它们在 55 摄氏度下固化一夜。在孵育结束时，从烤箱中取出重物和模具。接下来，在丙烯酸和固化的PDMS之间工作剃须刀片 打破密封 使用剃须刀或金属刮刀，小心地松开固化PDMS的侧面并将其从模具中取出。将 PDMS 设备包裹在铝箔中。用高压灭菌器胶带密封，并在 15 摄氏度和每平方英寸 15 磅的表压下通过干循环高压灭菌至少 121 分钟进行灭菌。要制备多孔装置，请将干珠浴培养箱预热至90摄氏度。打开高压灭菌的多孔装置的包装，并在装置的右上角切一个小凹口。将最多三个未包装的设备放入等离子清洁器中，孔朝上，然后运行等离子清洁器。通过用 70% 乙醇喷洒托盘内部并用任务擦拭将其擦干，对每个设备一个单孔聚苯乙烯托盘进行消毒。用戴手套的手从等离子清洁器中取出每个设备，并将其放入清洁的托盘中。然后将25毫升一次性移液器盆放入预热90摄氏度的珠浴培养箱中。每个要填充的设备取一管固化的LMNGM预混合物，并将其放入200毫升玻璃烧杯中。取下盖子和石蜡膜，然后微波，直到介质充分融化以倒入。将熔融的LMNGM预混合物倒入另一个无菌的200毫升烧杯中。如果一次填充多个设备，则可以在此烧杯中组合多个试管。将LMNGM预混合物再微波20秒。如果预混合物开始沸腾，请停止微波炉并让它沉降。从微波炉中取出熔融的LMNGM预混合物，并将其冷却至60摄氏度。将剩余的LMNGM成分添加到预混合混合物中，并将熔融的LMNGM倒入珠浴中的25毫升盆中。使用 200 微升多通道中继器移液器填充设备孔。将中继器设置为分配 14 微升的等分试样并安装五个移液器吸头。用熔融的LMNGM装载吸头，并将前14微升分配回装有LMNGM的盆中。快速但小心地将 14 微升分配到设备的内孔中，然后将剩余的 LMNGM 分配回盆中，因为最终等分试样通常小于 14 微升。接下来，设置中继器分配 15 微升的等分试样并填充最外层的孔环。使用 200 微升移液器，将 200 微升硫酸铜溶液分配到每个角落的设备护城河中。避免护城河过满，并确保硫酸铜不接触井的顶面。如果硫酸铜不容易流过整个护城河，请使用短铂丝镐来帮助打破张力并将硫酸铜拖过护城河。硫酸铜流过整个护城河后，使用200微升移液管或真空抽吸尽可能多地去除硫酸铜。使用挤压瓶，在设备和托盘壁之间的空间中添加水晶体。合上托盘盖，用一块封口膜包裹所有四个面。添加两个额外的封口膜片以完全密封板。使用中继移液器，用五微升浓缩细菌点缀每个孔。避免直接接触 LMNGM 表面。使用定制的干燥箱干燥设备，该干燥箱可以使用计算机机箱风扇和HEPA过滤器以最低的成本建造。在解剖范围内，检查孔以确保所有孔都已成功干燥。使用铂镐，从准备好的蜗虫板中转移动物，每孔添加一个蠕虫。将一滴准备好的洗涤剂溶液添加到托盘盖内侧，并用任务擦拭擦拭，直到洗涤剂溶液干燥。按照手稿中的说明，用三片石蜡像包裹托盘。daf-2（RNAi）的寿命延长被daf-16零突变阻断。在daf-2（RNAi）存在的情况下，健康寿命也减少了整个生命周期的三天滚动平均值，daf-16突变体和daf-2（RNAi）显示了个体动物的寿命和健康寿命的变化，以绝对值或总寿命的一小部分进行比较。个体动物在整个生命周期中的累积活动与寿命的相关性优于单个动物在整个生命周期中任何特定日期的活动。重要的是尽快填满井并添加蠕虫。否则，井可能会干涸，蠕虫将无法生存。该系统与自动成像兼容，可用于收集高通量数据，例如寿命、活动和体型或形状。

Research

JoVE Journal

Biology

Long-Term Culture and Monitoring of Isolated Caenorhabditis elegans on Solid Media in Multi-Well Devices

Measuring Caenorhabditis elegans Life Span on Solid Media

Measuring Caenorhabditis elegans Life Span on Solid Media

测量线虫固体介质的寿命

Aging is a degenerative process characterized by a progressive deterioration of cellular components and organelles resulting in mortality. The nematode ...

科研

生物学

Solid Plate-based Dietary Restriction in Caenorhabditis elegans

Solid Plate-based Dietary Restriction in Caenorhabditis elegans

实心板为基础的饮食限制在线虫</em

Reduction of food intake without malnutrition or starvation is known to increase lifespan and delay the onset of various age-related diseases in a wide ...

Time-lapse Microscopy of Early Embryogenesis in Caenorhabditis elegans

Time-lapse Microscopy of Early Embryogenesis in Caenorhabditis elegans

时间推移显微镜早期胚胎发育线虫

Caenorhabditis elegans has often been used as a model system in studies of early developmental processes.  The transparency of the 
embryos, the genetic ...

Long-term Culture of Human Breast Cancer Specimens and Their Analysis Using Optical Projection Tomography

长期培养的人类乳腺癌标本，并利用光学投影层析成像分析

Breast cancer is a leading cause of mortality in the Western world. It is well established that the spread of breast cancer, first locally and later ...

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

致命的甲壳类动物长期毒性试验卤虫franciscana</em

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish ...

Basic Caenorhabditis elegans Methods: Synchronization and Observation

Basic Caenorhabditis elegans Methods: Synchronization and Observation

基本的<em&gt;线虫</em&gt;方法：同步和观察

Research into the molecular and developmental biology of the nematode Caenorhabditis elegans was begun in the early seventies by Sydney Brenner and it has ...

Prostaglandin Extraction and Analysis in Caenorhabditis elegans

Prostaglandin Extraction and Analysis in Caenorhabditis elegans

前列腺素的提取与分析<em&gt;线虫</em

Caenorhabditis elegans is emerging as a powerful animal model to study the biology of lipids1-9. Prostaglandins are an important class of eicosanoids, ...

Culturing Caenorhabditis elegans in Axenic Liquid Media and Creation of Transgenic Worms by Microparticle Bombardment

Culturing Caenorhabditis elegans in Axenic Liquid Media and Creation of Transgenic Worms by Microparticle Bombardment

培养<em&gt;秀丽隐杆线虫</em &gt;在无菌液体介质与创造转基因蠕虫通过微粒轰击

In this protocol, we present the required materials, and the procedure for making modified&#160;C. elegans&#160;Habituation and Reproduction media ...

Isolation, Transfection, and Long-Term Culture of Adult Mouse and Rat Cardiomyocytes

成年小鼠和大鼠心肌细胞的隔离、转染和长期培养

Ex vivo culture of the adult mammalian cardiomyocytes (CMs) presents the most relevant experimental system for the in vitro study of cardiac biology. ...

Label-Free Imaging of Lipid Storage Dynamics in Caenorhabditis elegans using Stimulated Raman Scattering Microscopy

Label-Free Imaging of Lipid Storage Dynamics in Caenorhabditis elegans using Stimulated Raman Scattering Microscopy

使用刺激拉曼散射显微镜对 卡诺哈布迪炎电子人 中的脂质存储动力学进行无标签成像

Lipid metabolism is a fundamental physiological process necessary for cellular and organism health. Dysregulation of lipid metabolism often elicits ...

多孔设备固体培养基上分离秀 丽隐杆线虫 的长期培养和监测

多孔设备固体培养基上分离秀丽隐杆线虫的长期培养和监测