这项工作得到了加拿大自然科学与工程研究委员会发现授权RGPIN36481-08给William G. Bendena的支持。

1.实验生物的制备<ol><li> 在开始测定前24小时挑选每个菌株10只L4分期线虫，以确保生物体是年轻成年人。对于每个突变体或对照线虫，选择10个L4（对照为10个，测定为10个）。 <ol><li>维持使用标准方法26，在标准琼脂平板27 24小时与OP50 大肠杆菌接种L4生物体。如果在随后的洗涤步骤中生物体丢失，则通过增加起始样品量来补偿（ 即挑选20个蠕虫而不是10个）。 注意:行为是一种天生变异的表型。在三个不同的日子对每个菌株执行一式三份的方法。包括新的菌株的附加控制菌株和条件，如讨论部分所强调的。 </li></ol></li><li> 在测定之前，转移实验组织在没有细菌的琼脂平板上，允许线虫自由移动1分钟以除去多余的细菌。 <ol><li>如果实验生物在测定前24小时内发生污染状况，请丢弃。 </li></ol></li><li>移取1 mL M9到无菌板上，将蠕虫洗至微量离心管中。 </li><li>以3,000 xg离心1分钟。蠕虫应在管的底部形成颗粒。吸出M9溶液而不破坏蠕虫颗粒。向蠕虫颗粒中加入1 mL M9，将反应管与溶液混合。 </li><li> 重复步骤1.4三次。 <ol><li>如果过多的细菌最初用蠕虫转移，重复总共5次。不要将细菌转移到铜食品赛道板上。如果将食物转移到测定板上，则会干扰准确的数据收集。 </li></ol></li><li>最终洗涤后，吸出上清至10081; M的M和蠕虫颗粒残留。 注意:饥饿相关行为30分钟后变得明显。因此，一旦洗涤步骤完成，蠕虫应立即从溶液中转移。 </li></ol> 2.测定板的制备<ol><li> 在测定前两天准备标准的NGM琼脂平板。 <ol><li>如果将板保持在潮湿的环境中，在测定前3天制备琼脂板。或者，取下板的盖子3-6小时以确保适当的干燥（如果在无菌环境中）。 </li></ol></li><li>使用厚的永久标记，沿着外边缘在板的下侧形成一条线，另一条形成中线屏障（ 图1 ）。中线屏障应与板的每个边缘等距。使用标尺来确保精确的测量。这些生产线将作为传递细菌和铜溶液的指南。只要那个大肠杆菌在铜溶液前转移，这些线将作为指标。 </li><li> 种子板与50μL的OP50 大肠杆菌仅在铜屏障的一侧形成均匀的草坪（ 图2 ）。细菌浓度在测定中应保持一致;然而，随着浓度的轻微差异，观察到很小的变异性。 <ol><li>使用板下面的标记线，以确保细菌不与铜溶液接触。只要铜溶液排列在板的边缘并形成中线屏障，转移细菌使得铜溶液不会与食物源接触。 </li></ol></li><li>标记第二组平板，不转移OP50 大肠杆菌 （ 图2 ）。这些板块将用于评估阴性对照。这些盘子也应该有一个标记在板的一半上，以表示初始转移来源。 </li><li>允许细菌干燥，然后在37℃孵育平板过夜。确保转移到37°C的孵化器或房间时细菌斑块不会受到干扰。过度的干扰可能会改变食物贴片的位置或形状。 </li></ol>趋化因子测定<ol><li>在测定开始时间前新鲜制备0.5M硫酸铜（II）溶液。如果每个板使用125μL的溶液，则根据使用的测定板的数量（ 例如 5个测定板，625μL）来放大该体积。 </li><li>吸取100μL硫酸铜（II）溶液在琼脂边缘，以产生外部铜屏障。铭牌的下侧应作为指导。 </li><li> 移取25μL硫酸铜（II）溶液以产生中线屏障。 <ol><li>确保硫酸铜（II）溶液不接触细菌贴片。使用斑点技术可能会因为琼脂上的凹痕/划痕而影响运动。 </li></ol></li><li>让铜溶液干燥到板上。时间段可能因板材和实验室条件而异。转移后每5分钟检查干燥。 注意:铜溶液显示蓝色色调，易于识别。使用实验室组织轻轻地吸取板边缘附近的溶液，以辨别干燥。 </li><li> 将20μL蠕虫颗粒从管底移取到无细菌检测板的一半。 <ol><li>确保将10个蠕虫转移到测定板上。如果意外包含额外的蠕虫，请用卤代烃油取出，以确保没有细菌添加到板上。在测定期间，每个测定应具有一致数量的线虫。 注意:如果传染虫数太少，在测定过程中发生错误，增加初始样品量将减轻洗涤和转移期间的潜在损失。 </li></ol></li><li>用实验室组织从板上清除多余的M9。 M9不应与硫酸铜（II）溶液接触。 注意:确保蠕虫和琼脂表面在执行此步骤时不受影响。如果使用过度苛刻，实验室组织可能会对板的琼脂表面产生压痕，并可以去除蠕虫。应该丢弃通过 KimWipe意外删除的蠕虫。 </li><li> 一旦M9溶液被去除并且所有的蠕虫已经开始非液体运动模式，启动测定秒表。 <ol><li>最佳地，在一分钟内取出M9溶液。基本参数是识别正弦运动。蠕虫在液体中不同， 例如颠簸而不是正弦波。每次实验器官开始测定停止观察战争停止颠簸。 </li></ol></li><li> 每30分钟检查一次测定板。 <ol><li>对于具有细菌斑块的测定板，如果在4小时内到达食物贴片，则对生物有积极的作用。对于阴性对照板，如果他们已经越过障碍物，则对生物有积极的作用。 </li></ol></li></ol>

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我们没有什么可以披露的。

该测定设计修改了食物种族测定24，以包括铜溶液以产生厌恶的中线屏障并围绕板的边缘以防止线虫的丢失。测试生物体在四小时内穿过障碍物并达到食物补丁的能力。在npr-9（GF）的上下文中，我们利用该测定来评估饥饿条件如何影响厌食反应和食物检测。如果我们以前将npr-9（GF）表征为对食物和厌恶线索的反应性有缺陷，我们将多个环境线索与营养状况相结合，...

秀丽隐杆线虫已经被用作研究神经生物学数十年的模型，因为分析仅由302个神经元组成的神经系统的电路相对容易1 。只要生物依赖于环境线索的响应，大部分神经系统都致力于调节环境信号的整合2 。尽管神经系统的简单， C.线虫可以检测和响应各种环境信号，包括驱避剂3 ，引诱剂4 ，温度5 ，甚至湿度6 。正确地整合环境信号的故障一直挂在哺乳动物模型系统7-9一些行为障碍和神经退行性条件。有一系列可用的神经疾病模型<html> 在线虫中的 10种和线虫药物筛选11的发展，这种生物体已被证明是用于研究神经生物学的有用系统。鉴于线虫连接群1的可用性和线虫基因组12中几乎每个基因的突变，我们对线虫神经系统的理解以及我们自己的扩展都受到创造性地适合测定的设计的限制。 一些趋化实验已经开发在过去40年的线虫响应评估，以多样化的厌恶刺激3，4，13，14，15。初步实验涉及引入急性环境刺激，而单个蠕虫在琼脂板上漫游= "外部参照"&gt; 3，14，16。记录了对运动反应的立即改变。例如，可以将挥发性气味辛醇施用于头发，并在线虫鼻前摆放，以刺激野生型蠕虫17中向后运动的开始。更复杂的实验也已经发展到包含多个变量作为评估行为选择18的手段。该测定的变化需要使用铜溶液以产生厌恶的中线屏障4 。引诱剂，即二乙酰基，被放置在化学屏障的一侧，蠕虫从二乙酰基源转移出去。蠕虫对铜的厌恶反应有缺陷，马上越过障碍达到二乙酰，而野生型蠕虫最初被屏障排斥。当蠕虫第一次接近铜屏障时，响应得分没有长期观察。 蠕虫在经历饥饿状况后进行评估，对环境刺激的敏感性降低19 。当厌恶的化学辛醇在线虫鼻前飘荡时，野生型生物体在食物中会在3-5秒内刺激向后的运动。在这些生物从食物中取出10分钟后，它们的延迟反应为8 - 10秒20 。因此，随着饥饿的增加，线虫对有害环境信号的反应显示出减少的反应，因为对食物的追求变得对生存更为重要。相反，过表达神经肽受体9（ npr-9）的线虫对食物上或食物上的辛醇没有反应，并且表现出无法应对许多厌恶刺激21 。这些npr-9（GF）生物体也不会在食物存在的情况下调节其逆转频率，但可以反对刺激刺激的刺激表示他们能够向后运动21 。鉴于它们表现出异常下降的食物反转频率，但可以在食物21的情况下调节其行为，因此我们还评估了npr-9（LF）突变体。耦合引进急性外界刺激的蠕虫的营养状况阐明由食品相关的途径大致可以调节感官信号传导机制已经资助途径22，23。食物在线虫环境中的存在也被用于评估乙醇提取反应24 。在该实验中，将蠕虫在不同浓度的乙醇中孵育，然后置于具有被称为"食物种族测定"的食品的琼脂平板上。食物贴片放置在盘子的一个边缘，而线虫w远离食物来源。通过测量蠕虫到达食物块所需的时间来评估乙醇抽出。 这种基于营养的铜避免测定建立在食物种族测定的基础上，以整合额外的环境变量，即食物和铜，同时评估随时间推移的行为变化。这是整个线虫群落中常用协议的改编4 。该方案已被用于评估厌恶反应和食物在四小时内的检测21 。由于蠕虫在食物剥夺30分钟后出现饥饿行为25 ，我们还可以评估营养状况的变化如何影响环境反应。该测定的条件测量实验生物随时间改变对厌恶刺激的反应性，因此评估行为变化生物进入饥饿状态（并持续测量长时间的饥饿）。由于npr-9（GF）动物不会改变他们对食物或许多厌恶线索的行为，我们试图确定这些行为缺陷是否会在饥饿的背景下持续存在。最终，该测定设计已经被配制为特异性地评估npr-9（GF）突变体，但是可以进一步适用于表征新菌株。

<table><tbody><tr><td>M9 Solution [3 g KH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt;, 6 g Na&lt;sub&gt;2&lt;/sub&gt;HPO&lt;sub&gt;4&lt;/sub&gt;, 5 g NaCl, 1 ml 1 M MgSO&lt;sub&gt;4&lt;/sub&gt;, H&lt;sub&gt;2&lt;/sub&gt;O to 1 litre. Autoclave to sterilize before use.]</td><td></td><td></td><td>Produced in lab</td></tr><tr><td>Cupric Sulfate</td><td>Sigma</td><td>C-1297</td><td>Use water to appropriately suspend to a concentration of 0.5M</td></tr></tbody></table>

a caenorhabditis elegans nutritional-status based copper aversion assay

为了确保生存，生物必须能够避免不利的栖息地，同时确保一致的食物来源。 秀丽隐杆线虫在检测到各种环境刺激后改变其运动模式，并可以调整其对饥饿状况的行为反应套件。当从食物源中取出超过30分钟时，线虫通常表现出降低的反应反应。观察响应于不断变化的营养状况的行为变化可以提供对调节从喂养到饥饿状态的过渡的机制的了解。 我们已经开发了一种测定线虫测量过渡障碍（ 即铜），然后在长时间内达到食物来源的能力。该协议基于以前的工作，通过以允许随着生物向a转移的持续数据收集的方式整合多个变量来构建n越来越饿了。此外，该测定允许增加的样品量，使得可以同时评估较大的线虫种群。 检测或响应铜的能力的生物有机体立即穿越化学屏障，野生型线虫最初被排斥。随着野生型蠕虫越来越缺乏，它们开始越过障碍并达到食物来源。我们设计了这种检测方法来评估一种不能应对各种环境线索的突变体，包括食物感觉或厌恶性化学物质的检测。当通过该方案进行评估时，有缺陷的生物体立即越过障碍物，但也不能检测食物来源。因此，尽管临时达到食物来源，这些突变体重复地跨越化学屏障。该测定可以直接测试蠕虫群体，以评估与厌恶和饥饿相关的潜在途径缺陷。

我们利用野生型（N2）， npr-9（tm1652）和npr-9过表达菌株， 即 npr-9（GF） （IC836- npr-9 :: npr-9; sur-5 :: gfp; odr -1 :: rfp），以评估对饥饿和铜厌恶的反应。野生型生物体能够检测并响应于所述厌恶铜阻挡，而NPR-9（GF）的突变体不发起超过4小时测定21至铜厌恶响应。饥饿30分钟后，大约50-60％的野生型（N2）生物体穿过铜?...

Watch this Scientific Journal Video about A Caenorhabditis elegans Nutritional-status Based Copper Aversion Assay at JoVE.com

A Caenorhabditis elegans Nutritional-status Based Copper Aversion Assay

一个<em秀丽隐杆线虫</em基于营养状态的铜反转测定

在这里，我们提出了一种秀丽隐杆线虫特异性测定法，用于评估铜厌恶行为的变化和定位常见食物来源的能力，因为生物体从充足的营养状态进入饥饿的营养状态。

To ensure survival, organisms must be capable of avoiding unfavorable habitats while ensuring a consistent food source. Caenorhabditis elegans alter their ...

a-caenorhabditis-elegans-nutritional-status-based-copper-aversion

Research

JoVE Journal

Neuroscience

A Caenorhabditis elegans Nutritional-status Based Copper Aversion Assay

6.5K 次观看.  Queen's University. 在这里，我们提出了一种秀丽隐杆线虫特异性测定法，用于评估铜厌恶行为的变化和定位常见食物来源的能力，因为生物体从充足的营养状态进入饥饿的营养状态。 

视频: A Caenorhabditis elegans Nutritional-status Based Copper Aversion Assay

Automated Analysis of a Nematode Population-based Chemosensory Preference Assay

The nematode, Caenorhabditis elegans' compact nervous system of only 302 neurons underlies a diverse repertoire of behaviors. To facilitate the dissection of the neural circuits underlying these behaviors, the development of robust and reproducible behavioral assays is necessary. Previous C. elegans behavioral studies have used variations of a "drop test", a "chemotaxis assay", and a "retention assay" to investigate the response of C. elegans to soluble compounds. The method described in this article seeks to combine the complementary strengths of the three aforementioned assays. Briefly, a small circle in the middle of each assay plate is divided into four quadrants with the control and experimental solutions alternately placed. After the addition of the worms, the assay plates are loaded into a behavior chamber where microscope cameras record the worms' encounters with the treated regions. Automated video analysis is then performed and a preference index (PI) value for each video is generated. The video acquisition and automated analysis features of this method minimizes the experimenter's involvement and any associated errors. Furthermore, minute amounts of the experimental compound are used per assay and the behavior chamber's multi-camera setup increases experimental throughput. This method is particularly useful for conducting behavioral screens of genetic mutants and novel chemical compounds. However, this method is not appropriate for studying stimulus gradient navigation due to the close proximity of the control and experimental solution regions. It should also not be used when only a small population of worms is available. While suitable for assaying responses only to soluble compounds in its current form, this method can be easily modified to accommodate multimodal sensory interaction and optogenetic studies. This method can also be adapted to assay the chemosensory responses of other nematode species.

We present a behavior recording setup and protocol that enables automated analysis of the nematode, Caenorhabditis elegans' preference for soluble compounds in a population-based assay. This article describes the construction of a behavior chamber, the behavioral assay protocol, and video analysis software usage.

基于线虫人群的化学感应优选测定的自动分析

The nematode, Caenorhabditis elegans' compact nervous system of only 302 neurons underlies a diverse repertoire of behaviors. To facilitate the dissection ...

科研

行为学

A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans

Applying toxicity testing of chemicals in higher order organisms, such as mice or rats, is time-consuming and expensive, due to their long lifespan and maintenance issues. On the contrary, the nematode Caenorhabditis elegans (C. elegans) has advantages to make it an ideal choice for toxicity testing: a short lifespan, easy cultivation, and efficient reproduction. Here, we describe a protocol for the automatic phenotypic profiling of C. elegans in a 384-well plate. The nematode worms are cultured in a 384-well plate with liquid medium and chemical treatment, and videos are taken of each well to quantify the chemical influence on 33 worm features. Experimental results demonstrate that the quantified phenotype features can classify and predict the acute toxicity for different chemical compounds and establish a priority list for further traditional chemical toxicity assessment tests in a rodent model.

A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans

A quantitative method has been developed to identify and predict the acute toxicity of chemicals by automatically analyzing the phenotypic profiling of Caenorhabditis elegans. This protocol describes how to treat worms with chemicals in a 384-well plate, capture videos, and quantify toxicological related phenotypes.

一种用自动表型分析预测线虫化学毒性的高通量测定

Applying toxicity testing of chemicals in higher order organisms, such as mice or rats, is time-consuming and expensive, due to their long lifespan and ...

环境科学

测量 秀丽隐杆 线虫食物摄入量和寿命的高通量测定

Quantifying Food Intake in Caenorhabditis elegans by Measuring Bacterial Clearance

Feeding is an essential biological process for an organism&#39;s growth, reproduction, and survival. This assay aims to measure the food intake of Caenorhabditis elegans&#160;(C. elegans), an important parameter when studying the genetics of aging or metabolism. In most species, feeding is determined by measuring the difference between the amount of food provided and the amount left after a given time interval. The method presented here uses the same strategy to determine the feeding of C. elegans. It measures the amount of bacteria, the food source of C. elegans, cleared within 72 h. This method uses 96-well microtiter plates and has allowed the screening of hundreds of drugs for their ability to modulate food intake at a speed and depth not possible in other animal models. The strength of this assay is that it allows to measure feeding and lifespan simultaneously and directly measures the disappearance of food and, thus, is based on the same principles used for other organisms, facilitating species-to-species comparison.

作者聚焦：探索脂毒性与 HFpEF 之间的关系 

This protocol describes an assay to quantify Caenorhabditis elegans feeding rate based on measuring the clearance of bacteria in liquid culture.

通过测量细菌清除率来量化 秀丽隐杆线虫 的食物摄入量

Feeding is an essential biological process for an organism&#39;s growth, reproduction, and survival. This assay aims to measure the food intake of ...

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the development of more effective treatments is the relative lack of understanding of the molecular underpinnings of the effects of ethanol on behavior. The nematode, Caenorhabditis elegans (C.&#160;elegans), provides a useful model in which to generate and test hypotheses about the molecular effects of ethanol. Here, we describe an assay that has been developed and used to examine the roles of particular genes and environmental factors in behavioral responses to ethanol, in which locomotion is the behavioral output. Ethanol dose-dependently causes an acute depression of crawling on an agar surface. The effects are dynamic; animals exposed to a high concentration demonstrate an initial strong depression of crawling, referred to here as initial sensitivity, and then partially recover locomotion speed despite the continued presence of the drug. This ethanol-induced behavioral plasticity is referred to here as the development of acute functional tolerance. This assay has been used to demonstrate that these two phenotypes are distinct and genetically separable. The straightforward locomotion assay described here is suitable for examining the effects of both genetic and environmental manipulations on these acute behavioral responses to ethanol in C.&#160;elegans.

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

C. elegans is a useful model for studying the effects of ethanol on behavior. We present a behavioral assay that quantifies the effects of ethanol on the locomotion speed of crawling worms; both initial sensitivity and the development of acute functional tolerance to ethanol can be measured with this assay.

测量乙醇的影响上的步态速度的测定<em&gt;秀丽隐杆线虫</em

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the ...

神经科学

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

The folding and assembly of proteins is essential for protein function, the long-term health of the cell, and longevity of the organism. Historically, the function and regulation of protein folding was studied in vitro, in isolated tissue culture cells and in unicellular organisms. Recent studies have uncovered links between protein homeostasis (proteostasis), metabolism, development, aging, and temperature-sensing. These findings have led to the development of new tools for monitoring protein folding in the model metazoan organism Caenorhabditis elegans. In our laboratory, we combine behavioral assays, imaging and biochemical approaches using temperature-sensitive or naturally occurring metastable proteins as sensors of the folding environment to monitor protein misfolding. Behavioral assays that are associated with the misfolding of a specific protein provide a simple and powerful readout for protein folding, allowing for the fast screening of genes and conditions that modulate folding. Likewise, such misfolding can be associated with protein mislocalization in the cell. Monitoring protein localization can, therefore, highlight changes in cellular folding capacity occurring in different tissues, at various stages of development and in the face of changing conditions. Finally, using biochemical tools ex vivo, we can directly monitor protein stability and conformation. Thus, by combining behavioral assays, imaging and biochemical techniques, we are able to monitor protein misfolding at the resolution of the organism, the cell, and the protein, respectively.

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

To study the relationship between protein homeostasis, stress and aging, we monitored changes in protein folding by following protein dysfunction, protein localization in the cell and protein stability at the organismal, cellular and protein levels, using the genetically tractable metazoan Caenorhabditis elegans as a model system.

The folding and assembly of proteins is essential for protein function, the long-term health of the cell, and longevity of the organism. Historically, the ...

生物学

Caenorhabditis elegans as a Model System for Discovering Bioactive Compounds Against Polyglutamine-Mediated Neurotoxicity

Age-related misfolding and aggregation of pathogenic proteins are responsible for several neurodegenerative diseases. For example, Huntington&#39;s disease (HD) is principally driven by a CAG nucleotide repeat that encodes an expanded glutamine tract in huntingtin protein. Thus, the inhibition of polyglutamine (polyQ) aggregation and, in particular, aggregation-associated neurotoxicity is a useful strategy for the prevention of HD and other polyQ-associated conditions. This paper introduces generalized experimental protocols to assess the neuroprotective capacity of test compounds against HD using established polyQ transgenic Caenorhabditis elegans models. The AM141 strain is chosen for the polyQ aggregation assay as an age-associated phenotype of discrete fluorescent aggregates can be easily observed in its body wall at the adult stage due to muscle-specific expression of polyQ::YFP fusion proteins. In contrast, the HA759 model with strong expression of polyQ-expanded tracts in ASH neurons is used to examine neuronal death and chemoavoidance behavior. To comprehensively evaluate the neuroprotective capacity of target compounds, the above test results are ultimately presented as a radar chart with profiling of multiple phenotypes in a manner of direct comparison and direct viewing.

Caenorhabditis elegans as a Model System for Discovering Bioactive Compounds Against Polyglutamine-Mediated Neurotoxicity

Here, we present a protocol to assess the neuroprotective activities of test compounds in Caenorhabditis elegans, including polyglutamine aggregation, neuronal death, and chemoavoidance behavior, as well as an exemplary integration of multiple phenotypes.

秀丽隐杆线虫 作为发现生物活性化合物对抗聚谷氨酰胺介导的神经毒性的模型系统

Age-related misfolding and aggregation of pathogenic proteins are responsible for several neurodegenerative diseases. For example, Huntington&#39;s ...

Measuring Oxidative Stress Resistance of Caenorhabditis elegans in 96-well Microtiter Plates

Oxidative stress, which is the result of an imbalance between production and detoxification of reactive oxygen species, is a major contributor to chronic human disorders, including cardiovascular and neurodegenerative diseases, diabetes, aging, and cancer. Therefore, it is important to study oxidative stress not only in cell systems but also using whole organisms. C. elegans is an attractive model organism to study the genetics of oxidative stress signal transduction pathways, which are highly evolutionarily conserved.
Here, we provide a protocol to measure oxidative stress resistance in C. elegans in liquid. Briefly, ROS-inducing reagents such as paraquat (PQ) and H2O2 are dissolved in M9 buffer, and solutions are aliquoted in the wells of a 96 well microtiter plate. Synchronized L4/young adult C. elegans animals are transferred to the wells (5-8 animals/well) and survival is measured every hour until most worms are dead. When performing an oxidative stress resistance assay using a low concentration of stressors in plates, aging might influence the behavior of animals upon oxidative stress, which could lead to an incorrect interpretation of the data. However, in the assay described herein, this problem is unlikely to occur since only L4/young adult animals are being used. Moreover, this protocol is inexpensive and results are obtained in one day, which renders this technique attractive for genetic screens. Overall, this will help to understand oxidative stress signal transduction pathways, which could be translated into better characterization of oxidative stress-associated human disorders.

Measuring Oxidative Stress Resistance of Caenorhabditis elegans in 96-well Microtiter Plates

C. elegans is an attractive model organism to study signal transduction pathways involved in oxidative stress resistance. Here we provide a protocol to measure oxidative stress resistance of C. elegans animals in liquid phase, using several oxidizing agents in 96 well plates.

测量氧化应激性<em&gt;秀丽隐杆线虫</em&gt;在96孔微量滴定板

Oxidative stress, which is the result of an imbalance between production and detoxification of reactive oxygen species, is a major contributor to chronic ...

Long-Term Culture of Individual Caenorhabditis elegans on Solid Media for Longitudinal Fluorescence Monitoring and Aversive Interventions

Caenorhabditis elegans are widely used to study aging biology. The standard practice in C. elegans aging studies is to culture groups of worms on solid nematode growth media (NGM), allowing the efficient collection of population-level data for survival and other physiological phenotypes, and periodic sampling of subpopulations for fluorescent biomarker quantification. Limitations to this approach are the inability to (1) follow individual worms over time to develop age trajectories for phenotypes of interest and (2) monitor fluorescent biomarkers directly in the context of the culture environment. Alternative culture approaches use liquid culture or microfluidics to monitor individual animals over time, in some cases including fluorescence quantification, with the tradeoff that the culture environment is contextually distinct from solid NGM. The WorMotel is a previously described microfabricated multi-well device for culturing isolated worms on solid NGM. Each worm is maintained in a well containing solid NGM surrounded by a moat filled with copper sulfate, a contact repellent for C. elegans, allowing longitudinal monitoring of individual animals. We find copper sulfate insufficient to prevent worms from fleeing when subjected to aversive interventions common in aging research, including dietary restriction, pathogenic bacteria, and chemical agents that induce cellular stress. The multi-well devices are also molded from polydimethylsiloxane, which produces high background artifacts in fluorescence imaging. This protocol describes a new approach for culturing isolated roundworms on solid NGM using commercially available polystyrene microtrays, originally designed for human leukocyte antigen (HLA) typing, allowing the measurement of survival, physiological phenotypes, and fluorescence across the lifespan. A palmitic acid barrier prevents worms from fleeing, even in the presence of aversive conditions. Each plate can culture up to 96 animals and easily adapts to a variety of conditions, including dietary restriction, RNAi, and chemical additives, and is compatible with automated systems for collecting lifespan and activity data.

Long-Term Culture of Individual Caenorhabditis elegans on Solid Media for Longitudinal Fluorescence Monitoring and Aversive Interventions

Here, we present a protocol to culture isolated individual nematodes on solid media for lifelong physiological parameter tracking and fluorescence quantification. This culture system includes a palmitic acid barrier around single-worm wells to prevent animals from fleeing, allowing the use of aversive interventions, including pathogenic bacteria and chemical stressors.

在固体培养基上长期培养单个 秀丽隐杆线虫 ，用于纵向荧光监测和厌恶干预

Caenorhabditis elegans are widely used to study aging biology. The standard practice in C. elegans aging studies is to culture groups of worms on solid ...

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摘要

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此视频中的章节

基于线虫人群的化学感应优选测定的自动分析

一种用自动表型分析预测线虫化学毒性的高通量测定

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

测量乙醇的影响上的步态速度的测定

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

秀丽隐杆线虫作为发现生物活性化合物对抗聚谷氨酰胺介导的神经毒性的模型系统

测量氧化应激性

在固体培养基上长期培养单个秀丽隐杆线虫，用于纵向荧光监测和厌恶干预

一个

摘要

探索更多视频

此视频中的章节

基于线虫人群的化学感应优选测定的自动分析

一种用自动表型分析预测线虫化学毒性的高通量测定

通过测量细菌清除率来量化 秀丽隐杆线虫 的食物摄入量

通过测量细菌清除率来量化 秀丽隐杆线虫 的食物摄入量

通过测量细菌清除率来量化 秀丽隐杆线虫 的食物摄入量

测量乙醇的影响上的步态速度的测定

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

秀丽隐杆线虫 作为发现生物活性化合物对抗聚谷氨酰胺介导的神经毒性的模型系统

测量氧化应激性

在固体培养基上长期培养单个 秀丽隐杆线虫 ，用于纵向荧光监测和厌恶干预

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

通过测量细菌清除率来量化秀丽隐杆线虫的食物摄入量

秀丽隐杆线虫作为发现生物活性化合物对抗聚谷氨酰胺介导的神经毒性的模型系统

在固体培养基上长期培养单个秀丽隐杆线虫，用于纵向荧光监测和厌恶干预