作者承认哈蒙德湾生物站，大湖科学中心，美国地质调查局的贡献。我们也感谢梅林达帧博士在中心高级显微镜在密歇根州立大学在激光扫描共聚焦显微镜她的技术支持。这项研究是由赠款从大湖区渔业委员会YWCD和WML的支持。

1，溶液配制<ol><li>使1升10倍的0.1M磷酸盐缓冲盐水（PBS，pH值为7.4）:称取26.2克磷酸钠（一元），115克磷酸钠（二元），并87.66克NaCl等。溶于约800ml蒸馏水H 2 O，调节pH值，并把音量开到1升，用蒸馏水H 2 O <ol><li>使1升0.1M磷酸缓冲盐水（pH 7.4）:取100毫升10倍的PBS中，并加入900毫升蒸馏H 2 O。 </li><li>使1升0.1M磷酸缓冲液（pH 7.4）中含0.1％的Triton X-100:取百毫升10倍的PBS中，加入1 ml曲通X-100，和使体积达1升，用蒸馏水H 2 O。 </li></ol></li><li>使1微升10x 0.1M磷酸盐缓冲液（PB，pH值为7.4）:称取26.2克磷酸钠（一元）和115克磷酸钠（二元）。溶于约800ml蒸馏水H 2 O，调节pH值，并把音量开到1升，用蒸馏水H 2 O <ol><li>使1升0.1M磷酸缓冲液（pH7.4）:取100毫升10倍的PB，并添加900毫升蒸馏H 2 O </li><li>使1升0.1M磷酸缓冲液（pH7.4），0.1％的Triton X-100:取百毫升10倍的PB，加入1 ml曲通X-100，和使体积达1升，用蒸馏水H 2 O。 </li></ol></li><li>使400毫升水凝胶单体溶液（4％丙烯酰胺，0.25％双丙烯酰胺，4％的多聚甲醛，0.0075％过硫酸铵，在0.1M的PBS 0.0005％皂苷）:称取0.3克过硫酸铵和0.2克皂甙。添加210毫升蒸馏水H 2 O的加入40ml 40％的丙烯酰胺，将10毫升2％双丙烯酰胺加入40ml 10倍的PBS（pH 7.4）中，加入100毫升16％的多聚甲醛，拌匀。 </li><li>使4升清溶液（4％SDS在0.2M硼酸）:称取49.464克硼酸和160g的SDS。溶于约3.5升蒸馏水H 2 O，调节pH值至8.5，用NaOH，并把音量开到4 L。 </li><li>让1升80％的甘油溶液:测量800毫升甘油，加入200毫升蒸馏水H 2 O，拌匀。 </li></ol> 2，组织准备<ol><li>制备水凝胶的单体溶液和等份10毫升各15ml离心管中。 </li><li>解剖整个组织，并在4℃下在水凝胶2天固定确保该水凝胶溶液是至少10倍的组织的体积。 </li><li>使水凝胶固定的组织（在15ml离心管）到通风橱。 </li><li>聚合水凝胶固定的组织，加入5μLTEMED，拌匀，让坐在通风柜在室温下放置2-3小时。 </li><li>彻底去除多余的胶。注:额外的胶会阻碍染色在清算过程中和抗体的渗透。 </li><li>孵育固定的组织在10毫升的培养箱摇床设定在70rpm和50℃下进行数日清溶液直至组织变得透明。改变结算解决方案，至少每天一次，并从组织改变结算解决方案时，去除多余的胶。 </li><li>冲洗组织在10毫升0.1M的PB，0.1％的Triton X-100在70 R下午和37℃过夜。重复此步骤3次，并从该组织改变缓冲溶液时，除去过量的凝胶。 </li><li>孵育在PB中的第一抗体溶液澄清的组织用0.1％的Triton X-100在70rpm和37℃下进行2天。注意:请足够的初级抗体溶液淹没整个组织。 </li><li>漂洗在10毫升PB的组织用0.1％的Triton X-100，在70转和37℃培养过夜。重复此步骤3次，并从该组织改变缓冲溶液时，除去过量的凝胶。 </li><li>执行在一个黑暗的房间或昏暗光线下面的步骤。盖上铝箔样品，以防止照片褪色。 </li><li>孵育澄清组织与在PB中的荧光二抗溶液用0.1％的Triton X-100，并在70转和37℃下进行1天封闭血清。 </li><li>漂洗在10毫升PB的组织用0.1％的Triton X-100，在70转和37℃培养过夜。重复此步骤3次，去除多余的凝胶f改变缓冲溶液时，ROM中的组织。 </li><li>冲洗的组织在10毫升PBS中，在70转和37℃培养过夜。重复此步骤3次，并从该组织改变缓冲溶液时，除去过量的凝胶。 </li><li>孵育澄清组织在80％甘油中，在70转和37℃培养过夜。 </li><li>存储荧光染色的组织在4℃之前，激光共聚焦显微镜。 </li></ol>

<ol>
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作者什么都没有透露。

这个协议是由一个称为"净"12新方法，该交联完整的组织与聚丙烯酰胺以形成纳米多孔水凝胶，然后除掉组织的细胞膜，以实现光学透明性及透气性高分子修饰。 "清晰度"允许远程投射和神经系统局部电路接线完好组织成像。这种新方法可用于变态过程中可视化整个胆道系统在海七鳃鳗肝脏。这是生物学中的挑战，获得高分辨率的信息和保持从一个复杂的系统，例如在整个胆道系统或使用?...

海七鳃鳗通过三个不同的人生阶段发展1,2。幼虫海七鳃鳗（L）的花费大部分时间在洞穴作为底栖滤食性动物。在经历的外部形态和重组内脏3剧变7变质阶段，由此产生的少年（合资）输入期间，他们吸食血液和组织液从宿主鱼寄生阶段，增加体重的100倍以上。经过1.0〜1.5年在海洋或大湖主机喂鱼，大人在早春停止喂养和迁移到河流产卵，然后死亡1,2。 变态期间，海七鳃鳗肝脏失去胆囊和整个胆管树，进化的突变表型，模仿人类婴儿疾病胆道闭锁。婴儿胆道闭锁是一种罕见的小儿肝脏疾病严重的并发症4,5,6,7，8,9,10，但是胆道闭锁的发病机制和病因在很大程度上是未知4。出生后的两年内病人胆道闭锁死去，除非手术治疗（葛西程序）执行5。随后，这些患者需要丰富的临床管理及常肝移植6。胆道闭锁的发病机理许多理论被提出，如病毒感染，先天性畸形，自身免疫性疾病，并且毒性损伤。然而，每到胆道闭锁的发展所作的贡献仍未有定论7,8,9,10。 不像遭受病理胆道闭锁的婴儿，海七鳃鳗经受发育编程胆道闭锁没有广泛的坏死性炎症，纤维化或肝硬化10。在此过程中10，动物可能会受到短暂的胆汁淤积，但适应这种发展的条件通过从头合成和胆汁盐在发育胆道闭锁，除了已知的机制，如在肝11减少胆汁盐合成后肠的分泌。这在海七鳃鳗的发展过程提供了唯一的已知的机会检查胆道闭锁的进展。 一个名为"清晰度"新开发的方法，通过将完整的组织成一个光学透明的纳米多孔水凝胶12可实现高清晰度成像在复杂的哺乳动物神经系统。使用海七鳃鳗的肝脏和修改CLARITY协议，胆道病变的完好组织成像可以记录整个肝脏变态。

<table border="0" cellpadding="0" cellspacing="0" width="702">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">40% acrylamide&#160;</td>
			<td style="width:200px;">Bio-Rad</td>
			<td style="width:119px;">161-0140</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2% bis-acrylamide&#160;</td>
			<td>Bio-Rad</td>
			<td>161-0142</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TEMED</td>
			<td>Bio-Rad</td>
			<td>161-0800</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ammonium persulfate&#160;</td>
			<td>Sigma</td>
			<td>A3678-25G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">boric acid</td>
			<td>Sigma</td>
			<td>B7901-1KG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">saponin&#160;</td>
			<td>Sigma</td>
			<td>47036</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">sodium dodecyl sulfate&#160;</td>
			<td>Sigma</td>
			<td style="width:119px;">L337-500G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">sodium phosphate (monobasic)</td>
			<td>Sigma</td>
			<td>04269-1KG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">sodium phosphate (dibasic)</td>
			<td>Sigma</td>
			<td>S5136-1KG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Triton X-100</td>
			<td>Sigma</td>
			<td>X100-500ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">glycerol&#160;</td>
			<td>Sigma</td>
			<td>G9012-500ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">16% paraformaldehyde&#160;</td>
			<td>Electron Microscopy Sciences</td>
			<td>15710-S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NaOH pellets&#160;</td>
			<td>EMD</td>
			<td>SX0590-3</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">15 ml centrifuge tubes</td>
			<td style="width:200px;">Any brand</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">dissecting tools&#160;</td>
			<td style="width:200px;">Any brand</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

a new clarification method to visualize biliary degeneration during liver metamorphosis in sea lamprey (petromyzon marinus)

胆道闭锁是婴儿期一种罕见的疾病，估计有1 15,000频率在美国东南部，但在东亚国家较为常见，与1台5000报频率。虽然知道关于胆道闭锁的管理，其发病机制仍不清楚。海七鳃鳗（Petromyzon马里努斯 ）提供了一个独特的机会来审视胆管变性的机制和进展。海七鳃鳗发展经历三个不同的人生阶段:幼虫，寄生，和成人。在从幼虫寄生少年的过渡，海七鳃鳗经历蜕变与戏剧性的重组和重构的外部形态和内部器官。在肝脏中，整个胆道系统丢失，包括胆囊和胆管树。一个名为"清晰度"新开发的方法进行了修改，以澄清整个肝脏和结与变质海七鳃鳗肠道。该胆管变性过程是通过使用激光扫描共聚焦显微镜观察和海七鳃鳗变态过程中看出端倪。这种方法提供了一个强大的工具来研究胆道闭锁在一个独特的动物模型。

发生于肝胆系统的几个重要发展事件海七鳃鳗变态过程。胆管和胆囊发生凋亡和退化（ 图1）。与肝细胞标志物细胞角蛋白19（CK19，存在于两个胆管上皮细胞和肝细胞前和变态13之后），并使用共聚焦显微镜的抗凋亡BCL2基因标记物相结合的改性澄清法和染色法，在整个胆道系统沿Z轴（捕获图2）和3 -维结构被重建（ 图3）。激光扫描共聚焦显微镜通过?...

Watch this Scientific Journal Video about A New Clarification Method to Visualize Biliary Degeneration During Liver Metamorphosis in Sea Lamprey Petromyzon marinus at JoVE.com

A New Clarification Method to Visualize Biliary Degeneration During Liver Metamorphosis in Sea Lamprey Petromyzon marinus

海七鳃鳗失去变态在胆囊和胆管，类似于人类的胆道闭锁的过程。一种新的固定和澄清方法（CLARITY）进行了修改，使用可视化激光扫描共聚焦显微镜在整个胆道系统。这种方法提供了一个强大的工具来研究胆管变性。

一种新的澄清方法，以可视化胆道变性在肝癌变态海七鳃鳗（ Petromyzon马里努斯）

Biliary atresia is a rare disease of infancy, with an estimated 1 in 15,000 frequency in the southeast United States, but more common in East Asian ...

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A New Clarification Method to Visualize Biliary Degeneration During Liver Metamorphosis in Sea Lamprey (Petromyzon marinus)

13.7K 次观看.  Michigan State University. 海七鳃鳗失去变态在胆囊和胆管，类似于人类的胆道闭锁的过程。一种新的固定和澄清方法（CLARITY）进行了修改，使用可视化激光扫描共聚焦显微镜在整个胆道系统。这种方法提供了一个强大的工具来研究胆管变性。 

视频: A New Clarification Method to Visualize Biliary Degeneration During Liver Metamorphosis in Sea Lamprey Petromyzon marinus

Pyrosequencing: A Simple Method for Accurate Genotyping

Pharmacogenetic research benefits first-hand from the abundance of information provided by the completion of the Human Genome Project. With such a tremendous amount of data available comes an explosion of genotyping methods. Pyrosequencing(R) is one of the most thorough yet simple methods to date used to analyze polymorphisms. It also has the ability to identify tri-allelic, indels, short-repeat polymorphisms, along with determining allele percentages for methylation or pooled sample assessment. In addition, there is a standardized control sequence that provides internal quality control. This method has led to rapid and efficient single-nucleotide polymorphism evaluation including many clinically relevant polymorphisms. The technique and methodology of Pyrosequencing is explained.


Pyrosequencing(R) is one of the most thorough yet simple methods to date used to analyze polymorphisms. This method has led to rapid and efficient single-nucleotide polymorphism evaluation including many clinically relevant polymorphisms. The technique and methodology of Pyrosequencing is explained.

焦磷酸测序：一个准确的基因分型的简单方法

Pharmacogenetic research benefits first-hand from the abundance of information provided by the completion of the Human Genome Project. With such a ...

科研

生物学

Method for Culture of Early Chick Embryos ex vivo (New Culture)

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ideal tool for studying  the formation and patterning of brain, neural tube, somite and heart primordia. Applications of chick embryo culture include electroporation of DNA or RNA constructs in order to analyze gene function, grafts of growth factor coated beads such as FGFs and BMPs , as well as whole mount in situ hybridization and immunohistochemistry. This video demonstrates the different steps in chick embryo culture; First, the embryo is explanted in saline. Then, the embryo is centered on a glass ring. The membranes surrounding the embryo are lifted along the walls of the ring. The ring is then placed in a culture dish containing a pool of albumine. The culture dish is sealed and placed in a humid chamber, where the embryo is cultured for up to 24 hrs. Finally, the embryo is removed from the ring, fixed and processed for further applications. A troubleshooting guide is also presented.

Method for Culture of Early Chick Embryos ex vivo New Culture

This video demonstrates New culture, a method by which chick embryos are cultured outside the egg for up to 24 hr. This method enables one to study early development (primitive streak to 14 som.), a period corresponding to E7-9 in mouse. Applications of this technique include electroporation, in situ hybridization and immunohistochemistry.

文化早期鸡胚体外（新文化）的方法

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ...

Visually Mediated Odor Tracking During Flight in Drosophila

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual stabilization of upwind tracking directly aids in odor tracking. But do olfactory signals directly influence visual tracking behavior independently from wind cues? Additionally, recent advances in olfactory molecular genetics and neurophysiology have motivated novel quantitative behavioral analyses to assess the behavioral influence of (e.g.) genetically inactivating specific olfactory activation circuits. We modified a magnetic tether system originally devised for vision experiments by equipping the arena with narrow laminar flow odor plumes. Here we focus on experiments that can be performed after a fly is tethered and is able to navigate in the magnetic arena. We show how to acquire video images optimized for measuring body angle, how to judge stable odor tracking, and we illustrate two experiments to examine the influence of visual cues on odor tracking.

Here we describe how to optimize the acquired video image for an olfactory magnetic-tether (OMT) apparatus. We also describe two sample experimental protocols for studying visuo-olfactory fusion.

在果蝇的飞行过程中可视介导的气味追踪

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual ...

RNAi-mediated Gene Knockdown and In Vivo Diuresis Assay in Adult Female Aedes aegypti Mosquitoes

This video protocol demonstrates an effective technique to knockdown a particular gene in an insect and conduct a novel bioassay to measure excretion rate. This method can be used to obtain a better understanding of the process of diuresis in insects and is especially useful in the study of diuresis in blood-feeding arthropods that are able to take up huge amounts of liquid in a single blood meal. 
This RNAi-mediated gene knockdown combined with an in vivo diuresis assay was developed by the Hansen lab to study the effects of RNAi-mediated knockdown of aquaporin genes on Aedes aegypti mosquito diuresis1. 
The protocol is setup in two parts: the first demonstration illustrates how to construct a simple mosquito injection device and how to prepare and inject dsRNA into the thorax of mosquitoes for RNAi-mediated gene knockdown. The second demonstration illustrates how to determine excretion rates in mosquitoes using an in vivo bioassay.

RNAi-mediated Gene Knockdown and In Vivo Diuresis Assay in Adult Female Aedes aegypti Mosquitoes

In this protocol we combine RNAi-mediated gene silencing with an in vivo diuresis assay to study the effects knockdown of genes of interest has on mosquito fluid excretion.

RNAi介导的基因敲除和<em&gt;在体内</em在成年女性利尿含量<em&gt;埃及伊蚊</em&gt;蚊子

This video protocol demonstrates an effective technique to knockdown a particular gene in an insect and conduct a novel bioassay to measure excretion ...

Do-It-Yourself Device for Recovery of Cryopreserved Samples Accidentally Dropped into Cryogenic Storage Tanks

Liquid nitrogen is colorless, odorless, extremely cold (-196 &deg;C) liquid kept under pressure. It is commonly used as a cryogenic fluid for long term storage of biological materials such as blood, cells and tissues 1,2. The cryogenic nature of liquid nitrogen, while ideal for sample preservation, can cause rapid freezing of live tissues on contact - known as 'cryogenic burn'2, which may lead to severe frostbite in persons closely involved in storage and retrieval of samples from Dewars. Additionally, as liquid nitrogen evaporates it reduces the oxygen concentration in the air and might cause asphyxia, especially in confined spaces2.
In laboratories, biological samples are often stored in cryovials or cryoboxes stacked in stainless steel racks within the Dewar tanks1. These storage racks are provided with a long shaft to prevent boxes from slipping out from the racks and into the bottom of Dewars during routine handling. All too often, however, boxes or vials with precious samples slip out and sink to the bottom of liquid nitrogen filled tank. In such cases, samples could be tediously retrieved after transferring the liquid nitrogen into a spare container or discarding it. The boxes and vials can then be relatively safely recovered from emptied Dewar. However, the cryogenic nature of liquid nitrogen and its expansion rate makes sunken sample retrieval hazardous. It is commonly recommended by Safety Offices that sample retrieval be never carried out by a single person. Another alternative is to use commercially available cool grabbers or tongs to pull out the vials3. However, limited visibility within the dark liquid filled Dewars poses a major limitation in their use.
In this article, we describe the construction of a Cryotolerant DIY retrieval device, which makes sample retrieval from Dewar containing cryogenic fluids both safe and easy.

Here we present a low cost, durable cryotolerant device for sample retrieval from Dewar tanks filled with liquid nitrogen. The ease of construction and modular design of the device makes the process of sample retrieval from cryogenic tanks safe and easy.

做自己动手恢复冻存样品的设备不慎掉落到低温储罐

Liquid nitrogen is colorless, odorless, extremely cold (-196 &deg;C) liquid kept under pressure. It is commonly used as a cryogenic fluid for long term ...

Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization

Fluorescent in situ hybridization using DNA probes on 3-dimensionally preserved nuclei followed by 3D confocal microscopy (3D DNA FISH) represents the most direct way to visualize the location of gene loci, chromosomal sub-regions or entire territories in individual cells. This type of analysis provides insight into the global architecture of the nucleus as well as the behavior of specific genomic loci and regions within the nuclear space. Immunofluorescence, on the other hand, permits the detection of nuclear proteins (modified histones, histone variants and modifiers, transcription machinery and factors, nuclear sub-compartments, etc). The major challenge in combining immunofluorescence and 3D DNA FISH is, on the one hand to preserve the epitope detected by the antibody as well as the 3D architecture of the nucleus, and on the other hand, to allow the penetration of the DNA probe to detect gene loci or chromosome territories 1-5. Here we provide a protocol that combines visualization of chromatin modifications with genomic loci in 3D preserved nuclei.

Here we describe a protocol for simultaneous detection of histone modifications by immunofluorescence and DNA sequences by DNA FISH followed by 3D microscopy and analyses (3D immuno-DNA FISH).

结合免疫荧光和三维保存的间期核荧光原位杂交DNA研究在3D核组织的变化

Fluorescent in situ hybridization using DNA probes  on 3-dimensionally preserved nuclei followed by 3D confocal microscopy (3D DNA  FISH) represents the ...

High Throughput Microinjections of Sea Urchin Zygotes

Microinjection into cells and embryos is a common technique that is used to study a wide range of biological processes. In this method a small amount of treatment solution is loaded into a microinjection needle that is used to physically inject individual immobilized cells or embryos. Despite the need for initial training to perform this procedure for high-throughput delivery, microinjection offers maximum efficiency and reproducible delivery of a wide variety of treatment solutions (including complex mixtures of samples) into cells, eggs or embryos. Applications to microinjections include delivery of DNA constructs, mRNAs, recombinant proteins, gain of function, and loss of function reagents. Fluorescent or colorimetric dye is added to the injected solution to enable instant visualization of efficient delivery as well as a tool for reliable normalization of the amount of the delivered solution. The described method enables microinjection of 100-400 sea urchin zygotes within 10-15 min.

Microinjection is a common technique used to deliver DNA constructs, mRNAs, morpholino antisense oligonucleotides or other treatments into eggs, embryos, and cells of various species.

海胆受精卵的高通量显微注射

Microinjection into cells and embryos is a common technique that is used to study a wide range of biological processes. In this method a small amount of ...

A Method for Extracting Pigments from Squid Doryteuthis pealeii

Cephalopods can undergo rapid and adaptive changes in dermal coloration for sensing, communication, defense, and reproduction purposes. These capabilities are supported in part by the areal expansion and retraction of pigmented organs known as chromatophores. While it is known that the chromatophores contain a tethered network of pigmented granules, their structure-function properties have not been fully detailed. We describe a method for isolating the nanostructured granules in squid Doryteuthis pealeii chromatophores and demonstrate how their associated pigments can be extracted in acidic solvents. To accomplish this, the chromatophore containing dermal layer is first manually isolated using a superficial dissection, and the pigment granules are removed using sonication, centrifugation, and washing cycles. Pigments confined within the purified granules are then extracted via acidic methanol solutions, leaving nanostructures with smaller diameters that are void of visible color. This extraction procedure produces a 58% yield of soluble pigments isolated from granules. Using this method, the composition of the chromatophore pigments can be determined and used to provide insight into the mechanism of adaptive coloration in cephalopods.

A Method for Extracting Pigments from Squid Doryteuthis pealeii

A protocol for the extraction of pigments from the nanostructured granules in squid Doryteuthis pealeii chromatophores is presented.

一种从鱿鱼提取色素的方法<em&gt; Doryteuthis pealeii</em

Cephalopods can undergo rapid and adaptive changes in dermal coloration for sensing, communication, defense, and reproduction purposes. These capabilities ...

Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus

This work describes the whole-mount immunohistochemistry staining method in detail, using neurofilament protein antibody to label the innervation of the biliary tract in Suncus murinus (S. murinus&#160;). First, the specimen was dissected from S. murinus and fixed in 4% paraformaldehyde (PFA). Second, an enzymatic treatment and potential endogenous peroxidase inactivation were performed. The specimen was then exposed to the primary antibody, anti-neurofilament protein antibody, for 3-6 days. It was then incubated with the secondary antibody conjugated with horseradish peroxidase. The color reaction was revealed by reacting the specimen with a 3,3&#39;-diaminobenzidine (DAB) substrate. This method can be applied to analyze the innervation of all visceral organs. Furthermore, this protocol can also be adapted to test other neuronal antibodies, but optimization of the antibodies should be done first. This method was originally introduced by Kuratani and Tanaka1,2,3.

Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus

A whole-mount immunohistochemical approach, to visualize neurofilament protein expression in the extrahepatic biliary tract in Suncus murinus. is presented here. This protocol can be used to analyze the innervation of all visceral organs in S. murinus or other species.

使用全身免疫组织化学方法研究胆道的神经支配<em&gt; Suncus murinus</em

This work describes the whole-mount immunohistochemistry staining method in detail, using neurofilament protein antibody to label the innervation of the ...

Broth Microdilution In Vitro Screening: An Easy and Fast Method to Detect New Antifungal Compounds

Fungal infections have become an important medical condition in the last decades, but the number of available antifungal drugs is limited. In this scenario, the search for new antifungal drugs is necessary. The protocol reported here details a method to screen peptides for their antifungal properties. It is based on the broth microdilution susceptibility test from the Clinical and Laboratory Standards Institute (CLSI) M27-A3 guidelines with modifications to suit the research of antimicrobial peptides as potential new antifungals. This protocol describes a functional assay to evaluate the activity of antifungal compounds and may be easily modified to suit any particular class of molecules under investigation. Since the assays are performed in 96-well plates using small volumes, a large-scale screening can be completed in a short amount of time, especially if carried out in an automation setting. This procedure illustrates how a standardized and adjustable clinical protocol can help the bench-work pursuit of new molecules to improve the therapy of fungal diseases.

Broth Microdilution In Vitro Screening: An Easy and Fast Method to Detect New Antifungal Compounds

An easy and adaptable broth microdilution method for screening antifungal compounds and extracts.

一种新的澄清方法，以可视化胆道变性在肝癌变态海七鳃鳗（ Petromyzon马里努斯）

摘要

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此视频中的章节

焦磷酸测序：一个准确的基因分型的简单方法

文化早期鸡胚体外（新文化）的方法

在果蝇的飞行过程中可视介导的气味追踪

RNAi介导的基因敲除和

做自己动手恢复冻存样品的设备不慎掉落到低温储罐

结合免疫荧光和三维保存的间期核荧光原位杂交DNA研究在3D核组织的变化

海胆受精卵的高通量显微注射

一种从鱿鱼提取色素的方法

使用全身免疫组织化学方法研究胆道的神经支配

肉汤稀释体外筛选: 一种简便快速检测新抗真菌化合物的方法