作者想感谢他的技术支持Ikiel Ptito。弗兰克欧文，罗伯塔Palmour和行为科学基金会设在圣基茨实验室，西印度群岛，为他们继续支持我们的灵长类动物的工作人员，我们非常感谢。

第1部分：前处理的组织<ol><li>组织应多聚甲醛，戊二醛，或福尔马林灌注。这可以通过标准transcardial，通常用于收获其他器官灌注。建议后不久牺牲的眼睛被注入固定液，只是镜头下的固定液中。 </li><li>在本研究的主题是深刻的盐酸氯胺酮（10毫克/公斤，IM）与戊巴比妥钠（25毫克/公斤，四）过量安乐死，镇静剂和用0.1 M PBS灌注，直到完全exsanguinated transcardially人。其次是在PBS的4％多聚甲醛为5分钟（约1升）的解决方案。 </li></ol>第2部分：眼球去除眶腔<ol><li>更容易获得眼球，建议先取出大脑。一旦大脑已被删除的薄壁轨道骨是显而易见的。使用骨rongeurs慢慢芯片了墙上的轨道。用手术刀切去眼部肌肉和结缔组织从眼球中删除。小心地剪断视神经，这可以用在电子显微镜研究。眼球，现在应该被释放的眶腔。 </li></ol>第3部分：视网膜解剖从眼罩<ol><li>放入一个培养皿中，用PBS保持干燥视网膜的眼睛。在解剖显微镜或表安装在解剖放大光的立场是有益的，但​​不是必须的。取出一双弹簧剪刀锯齿缘水平密切切割巩膜角膜周边角膜和钳取出镜头。 </li><li>使用画笔，镊子，和弹簧剪刀去除巩膜视网膜。这是脱离巩膜视网膜，然后用剪刀切割巩膜，。我们必须开展以小增量这个过程中，以免损伤或撕裂的视网膜组织。巩膜是不容易分开的残余视神经这么仔细地减少视神经周围巩膜。 </li><li>此时视网膜保留其弯曲的形状，需要进行采样夷为平地。在扁平化的视网膜，玻璃体，里面有果冻的一致性，可以去掉一笔。要拉平到幻灯片上的视网膜，用手术刀刀片的径向削减。残留的玻璃体，现在可以删除与普通过滤纸和画笔。在这一点上，所以它必须要轻柔取出时玻璃体视网膜神经节细胞层暴露。如果视神经仍然附着在视盘，删除不使用手术刀刀片和一对弹簧剪刀翻录的视网膜视神经。这是一个平坦的装载视网膜中央凹（图1）和应作为一个黑暗的补丁在视盘颞/腹侧方向明显。 </li></ol>第4部分：采样<ol><li>有很多的选项，抽样视网膜。在这里，我们将描述flatmount准备和isodentric采样。对于这两个程序flatmount视网膜与视神经纤维层离幻灯片。如果意图是在flatmount准备检查视网膜神经节细胞层，这是必要的删除或漂白色素上皮。为了消除色素上皮使用画笔轻轻去除这层，这往往会损坏感光层。漂白涉及视网膜浸泡在高锰酸钾溶液（0.25％），1小时，用蒸馏水洗涤，并结算草酸（5％）5 分钟 2 。 </li><li>如果目的是研究整个视网膜细胞的分布和每一层的形态，那么我们建议视网膜等距抽样。视网膜应flatmounted幻灯片上，用PBS，并保持湿润。对于等距抽样切小块组织从视盘鼻，颞部，上下方向（图1）等距。这是没有必要的准备漂白色素上皮的。这些作品现在可以在冠状面切片与一个恒温器，vibratome，或根据所研究的问题ultramicrotome。低温恒温切片，件应在30％蔗糖过夜和cryoprotected，在安装介质上干冰中冷冻。另外，样品可以嵌入琼脂，在一个vibratome切片。低温恒温器和vibratome准备工作，将可靠的产量高达4米薄的部分如果需要较薄（例如电子显微镜准备），这是必要的，准备ultramicrotome样品。要做到这一点，部分应在四氧化锇后固定的通风柜下为1小时。其次是在梯度乙醇系列脱水（50，70，95，95，100，100％）和100％的环氧丙烷。组织然后嵌入在EPON（EMBED - 812嵌入套件）。 </li></ol>第5部分：代表性的成果： 在我们的实验室，我们进行常规免疫组织化学cryosectioned视网膜（图2）。在这种情况下，我们感兴趣的是在灵长类动物视网膜的大麻素受体（CB1）等密度。我们还研究灵长类动物的视觉系统的产前乙醇暴露的影响。为此，我们有兴趣在黄斑中心凹，并在周边视网膜细胞密度和层厚度。而要做到这一点，我们在EPON中嵌入视网膜件，切片在700nm的上ultramicrotome，和染色用1％甲苯胺蓝（图3）。 <img src="/files/ftp_upload/1261/fig1.jpg" alt="图1" /> 图1 Flatmount视网膜。径向削减用于扁平化的视网膜。 <img src="/files/ftp_upload/1261/fig2.jpg" alt="图2" /> 图2免疫染色。周边视网膜Cryosection免疫染色为CB1视网膜神经节细胞很大程度上与一些内部和外部的核层的标签标示。本节的厚度为14米，在幻灯片上染色。 RG - 视网膜神经节层，IP - 内丛状层; - 内核层; OP - 外丛状层; - 外核层。 <img src="/files/ftp_upload/1261/fig3.jpg" alt="图3" /> 图3通过嵌入在EPON和切片在700nm的ultramicrotrome中心凹中心凹。节。感光层（PR）是用于调整的部分。请注意，显示在冠状面的一个适当的角度可以识别的整个范围的光感受器。密度测量在300米，500米和800米Bioquant成像系统的中心凹坑中心。 RG - 视网膜神经节层，IP - 内丛状层; - 内核层; OP - 外丛状层; - 外核层;比例尺= 50米

<ol>
	<li>Hendrickson, A., Kupfer, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+histogenesis+of+the+fovea+in+the+macaque+monkey.">The histogenesis of the fovea in the macaque monkey.</a> Invest Ophthalmol Vis Sci. 15, 746-756 (1976).</li><li>Herbin, M., Boire, D., Ptito, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Size+and+distribution+of+retinal+ganglion+cells+in+the+St.+Kitts+green+monkey+(Cercopithecus+aethiops+sabeus).">Size and distribution of retinal ganglion cells in the St. Kitts green monkey (Cercopithecus aethiops sabeus).</a> J Comp Neurol. 383, 459-472 (1997).</li><li>Stone, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Wholemount Handbook. , (1981).</li><li>Herbin, M., Boire, D., Theoret, H., Ptito, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transneuronal+degeneration+of+retinal+ganglion+cells+in+early+hemispherectomized+monkeys.">Transneuronal degeneration of retinal ganglion cells in early hemispherectomized monkeys.</a> Neuroreport. 10, 1447-1452 (1999).</li><li>Krebs, W., Krebs, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Primate Retina and Choroid Atlas of Fine Structure in Man and Monkey. , (1991).</li><li>Pow, D., Sullivan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+kinesis,+neurite+sprouting+and+abnormal+axonal+projections+of+cone+photoreceptors+in+the+aged+and+AMD-afflicted+human+retina.">Nuclear kinesis, neurite sprouting and abnormal axonal projections of cone photoreceptors in the aged and AMD-afflicted human retina.</a> Exp Eye Res. 84, 850-857 (2007).</li></ol>

准备允许作为wholemount视网膜的神经节细胞层，或视网膜血管 3内皮细胞的地形和空间分布的分析。在灵长类动物的视网膜边缘的细胞密度的量化很容易完成。然而，在perifoveal和中心凹区域，堆积在神经节细胞层的多层次的阻碍量化。为了规避这种潜在的偏见，黄斑中心凹和perifoveal地区可以解剖从wholemount准备，嵌在EPON，并连续切片使用ultramicrotome获得半薄切片，在冠状面 2,4 。有w...

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the gateway to the brain: dissecting the primate eye

在人类的视觉系统被认为是世界的门户和过多的感官，知觉和认知过程中起着主要作用。因此，并不令人惊讶的视觉质量与生活质量。尽管广泛的临床和基础研究周围的视觉障碍的原因，许多形式如色素性视网膜炎和黄斑变性，视力障碍，缺乏有效的治疗方法。非人类灵长类动物的最接近人类的眼球发育一般特征。他们不仅有类似的血管解剖，但其他哺乳动物，包括灵长类动物有独特的特征，在一个区域专门用于高视力，黄斑中心凹视网膜颞<sup&gt; 1</sup&gt;。在这里，我们描述了一个解剖的灵长类动物的视网膜提供视网膜组织学，免疫组织化学，激光捕获显微切割技术，以及光学和电子显微镜的组织，一般技术。随着使用的非人类灵长类动物作为一个翻译模型的扩展，我们希望的是更好地理解视网膜将提供有效的方法以减轻或扭转受影响的个人的生活质量产生负面影响视觉障碍的见解。

Watch this Scientific Journal Video about The Gateway to the Brain: Dissecting the Primate Eye at JoVE.com

The Gateway to the Brain: Dissecting the Primate Eye

非人类灵长类动物是我们的发育和衰老的理解的重要翻译物种。灵长类动物视网膜的解剖组织可能为在人体正常和病理条件下的重要见解。

到大脑的门户：剖析灵长类动物的眼睛

The visual system in humans is considered the  gateway  to the world and plays a principal role in the plethora of sensory, perceptual and cognitive ...

the-gateway-to-the-brain-dissecting-the-primate-eye

Research

JoVE Journal

Biology

14.1K 次观看.  University of Montreal. 非人类灵长类动物是我们的发育和衰老的理解的重要翻译物种。灵长类动物视网膜的解剖组织可能为在人体正常和病理条件下的重要见解。

视频: The Gateway to the Brain: Dissecting the Primate Eye

Dissecting and Recording from The C. Elegans Neuromuscular Junction

Neurotransmission is the process by which neurons transfer information via chemical signals to their post-synaptic targets, on a rapid time scale. This complex process requires the coordinated activity of many pre- and post-synaptic proteins to ensure appropriate synaptic connectivity, conduction of electrical signals, targeting and priming of secretory vesicles, calcium sensing, vesicle fusion, localization and function of postsynaptic receptors and finally, recycling mechanisms. As neuroscientists it is our goal to elucidate which proteins function in each of these steps and understand their mechanisms of action. Electrophysiological recordings from synapses provide a quantifiable read out of the underlying electrical events that occur during synaptic transmission. By combining this technique with the powerful array of molecular and genetic tools available to manipulate synaptic proteins in C. elegans, we can analyze the resulting functional changes in synaptic transmission. 
The C. elegans NMJs formed between motor neurons and body wall muscles control locomotion, therefore, mutants with uncoordinated locomotory phenotypes (known as unc s) often perturb synaptic transmission at these synapses 1. Since unc mutants are maintained on a rich supply of a bacterial food source, they remain viable as long as they retain some pharyngeal pumping ability to ingest food. This, together with the fact that C. elegans exist as hermaphrodites, allows them to pass on mutant progeny without the need for elaborate mating behaviors. These attributes, coupled with our recent ability to record from the worms NMJs 2,3,7 make this an excellent model organism in which to address precisely how unc mutants impact neurotransmission. 
The dissection method involves immobilizing adult worms using a cyanoacrylic glue in order to make an incision in the worm cuticle exposing the NMJs. Since C. elegans adults are only 1 mm in length the dissection is performed with the use of a dissecting microscope and requires excellent hand-eye coordination. NMJ recordings are made by whole-cell voltage clamping individual body wall muscle cells and neurotransmitter release can be evoked using a variety of stimulation protocols including electrical stimulation, light-activated channel-rhodopsin-mediated depolarization 4 and hyperosmotic saline, all of which will be briefly described.

Application of electrophysiology to accessible synapses provides a quantifiable measure of synaptic activity, useful in analyzing synaptic mutants. This article describes a dissection method used to expose the neuromuscular junctions (NMJ) of Caenorhabditis elegans (C. elegans) and briefly discusses some of the uses to which this preparation can be applied.

线虫神经肌肉交界处的解剖和录制

Neurotransmission is the process by which neurons transfer information via chemical signals to their post-synaptic targets, on a rapid time scale.   This ...

科研

生物学

Lens Transplantation in Zebrafish and its Application in the Analysis of Eye Mutants

The lens plays an important role in the development of the optic cup[1,2]. Using the zebrafish as a model organism, questions regarding lens development can be addressed. The zebrafish is useful for genetic studies due to several advantageous characteristics, including small size, high fecundity, short lifecycle, and ease of care. Lens development occurs rapidly in zebrafish. By 72 hpf, the zebrafish lens is functionally mature [3]. Abundant genetic and molecular resources are available to support research in zebrafish. In addition, the similarity of the zebrafish eye to those of other vertebrates provides basis for its use as an excellent animal model of human defects[4-7]. Several zebrafish mutants exhibit lens abnormalities, including high levels of cell death, which in some cases leads to a complete degeneration of lens tissues [8]. To determine whether lens abnormalities are due to intrinsic causes or to defective interactions with the surrounding tissues, transplantation of a mutant lens into a wild-type eye is performed. Using fire-polished metal needles, mutant or wild-type lenses are carefully dissected from the donor animal, and transferred into the host. To distinguish wild-type and mutant tissues, a transgenic line is used as the donor. This line expresses membrane-bound GFP in all tissues, including the lens. This transplantation technique is an essential tool in the studies of zebrafish lens mutants.

Lens development involves interactions with other tissues. Several zebrafish eye mutants are characterized by an abnormally small lens size. Here we demonstrate a lens transplantation experiment to determine whether this phenotype is due to intrinsic causes or defective interactions with tissues that surround the lens.

在斑马鱼和镜头移植在眼突变分析中的应用

The lens plays an important role in the development of the optic cup[1,2]. Using the zebrafish as a model organism, questions regarding lens development ...

Dissecting the Non-human Primate Brain in Stereotaxic Space

The use of non-human primates provides an excellent translational model for our understanding of developmental and aging processes in humans1-6. In addition, the use of non-human primates has recently afforded the opportunity to naturally model complex psychiatric disorders such as alcohol abuse7. Here we describe a technique for blocking the brain in the coronal plane of the vervet monkey (Chlorocebus aethiops sabeus) in the intact skull in stereotaxic space. The method described here provides a standard plane of section between blocks and subjects and minimizes partial sections between blocks. Sectioning a block of tissue in the coronal plane also facilitates the delineation of an area of interest. This method provides manageable sized blocks since a single hemisphere of the vervet monkey yields more than 1200 sections when slicing at 50&mu;m. Furthermore by blocking the brain into 1cm blocks, it facilitates penetration of sucrose for cyroprotection and allows the block to be sliced on a standard cryostat.

The non-human primate is an important translational species for our understanding of the normal processing of the brain. The anatomical organization of the primate brain can provide important insights into normal and pathological conditions in humans.

剖析在非人类灵长类动物脑立体定向空间

The use of non-human primates provides an excellent translational model for our understanding of developmental and aging processes in humans1-6. In ...

Knowing What Counts: Unbiased Stereology in the Non-human Primate Brain

The non-human primate is an important translational species for understanding the normal function and disease processes of the human brain. Unbiased stereology, the method accepted as state-of-the-art for quantification of biological objects in tissue sections2, generates reliable structural data for biological features in the mammalian brain3. The key components of the approach are unbiased (systematic-random) sampling of anatomically defined structures (reference spaces), combined with quantification of cell numbers and size, fiber and capillary lengths, surface areas, regional volumes and spatial distributions of biological objects within the reference space4. Among the advantages of these stereological approaches over previous methods is the avoidance of all known sources of systematic (non-random) error arising from faulty assumptions and non-verifiable models. This study documents a biological application of computerized stereology to estimate the total neuronal population in the frontal cortex of the vervet monkey brain (Chlorocebus aethiops sabeus), with assistance from two commercially available stereology programs, BioQuant Life Sciences and Stereologer (Figure 1). In addition to contrast and comparison of results from both the BioQuant and Stereologer systems, this study provides a detailed protocol for the Stereologer system.

The anatomical organization of the primate brain can provide important insights into normal and pathological conditions in humans. Unbiased stereology is a method for accurately and efficiently estimating the total neuron number (or other cell type) in a given reference space1.

知道重要的：无偏体视学在非人类灵长类动物的脑

The non-human primate is an important translational species for understanding the normal function and disease processes of the human brain. Unbiased ...

A Novel RFP Reporter to Aid in the Visualization of the Eye Imaginal Disc in Drosophila

The Drosophila eye is a powerful model system for studying areas such as neurogenesis, signal transduction and neurodegeneration. Many of the discoveries made using this system have taken advantage of the spatiotemporal nature of photoreceptor differentiation in the developing eye imaginal disc. To use this system it is first necessary for the researcher to learn to identify and dissect the eye disc. We describe a novel RFP reporter to aid in the identification of the eye disc and the visualization of specific cell types in the developing eye. We detail a methodology for dissection of the eye imaginal disc from third instar larvae and describe how the eye-RFP reporter can aid in this dissection. This eye-RFP reporter is only expressed in the eye and can be visualized using fluorescence microscopy either in live tissue or after fixation without the need for signal amplification. We also show how this reporter can be used to identify specific cells types within the eye disc. This protocol and the use of the eye-RFP reporter will aid researchers using the Drosophila eye to address fundamentally important biological questions.

A Novel RFP Reporter to Aid in the Visualization of the Eye Imaginal Disc in Drosophila

We describe a novel red fluorescent protein (RFP) reporter that is expressed specifically in the Drosophila eye. We detail a methodology for dissection of the eye imaginal disc and how this reporter can be used to aid in the dissection and identification of specific cell types in the developing eye.

一种新型的RFP的记者，以帮助在眼成虫光盘的可视化果蝇</em

The Drosophila eye is a powerful model system for studying areas such as neurogenesis, signal transduction and neurodegeneration. Many of the discoveries ...

Sampling Blood from the Lateral Tail Vein of the Rat

Blood samples are commonly obtained in many experimental contexts to measure targets of interest, including hormones, immune factors, growth factors, proteins, and glucose, yet the composition of the blood is dynamically regulated and easily perturbed. One factor that can change the blood composition is the stress response triggered by the sampling procedure, which can contribute to variability in the measures of interest. Here we describe a procedure for blood sampling from the lateral tail vein in the rat. This procedure offers significant advantages over other more commonly used techniques. It permits rapid sampling with minimal pain or invasiveness, without anesthesia or analgesia. Additionally, it can be used to obtain large volume samples (upwards of 1 ml in some rats), and it may be used repeatedly across experimental days. By minimizing the stress response and pain resulting from blood sampling, measures can more accurately reflect the true basal state of the animal, with minimal influence from the sampling procedure itself.

Blood samples are useful for assessing biomarkers of physiological states or disease in vivo. Here we describe the methodology to sample blood from the lateral tail vein in the rat. This method provides rapid samples with minimal pain and invasiveness.

从大鼠的侧尾静脉血液取样

Blood samples are commonly obtained in many experimental contexts to measure targets of interest, including hormones, immune factors, growth factors, ...

Adipo-Clear: A Tissue Clearing Method for Three-Dimensional Imaging of Adipose Tissue

Adipose tissue plays a central role in energy homeostasis and thermoregulation. It is composed of different types of adipocytes, as well as adipocyte precursors, immune cells, fibroblasts, blood vessels, and nerve projections. Although the molecular control of cell type specification and how these cells interact have been increasingly delineated, a more comprehensive understanding of these adipose-resident cells can be achieved by visualizing their distribution and architecture throughout the whole tissue. Existing immunohistochemistry and immunofluorescence approaches to analyze adipose histology rely on thin paraffin-embedded sections. However, thin sections capture only a small portion of tissue; as a result, the conclusions can be biased by what portion of tissue is analyzed. We have therefore developed an adipose tissue clearing technique, Adipo-Clear, to permit comprehensive three-dimensional visualization of molecular and cellular patterns in whole adipose tissues. Adipo-Clear was adapted from iDISCO/iDISCO+, with specific modifications made to completely remove the lipid stored in the tissue while preserving native tissue morphology. In combination with light-sheet fluorescence microscopy, we demonstrate here the use of the Adipo-Clear method to obtain high-resolution volumetric images&#160;of an entire adipose tissue.

Due to the high lipid content, adipose tissue has been challenging to visualize using traditional histological methods. Adipo-Clear is a tissue clearing technique that allows robust labeling and high-resolution volumetric fluorescent imaging of adipose tissue. Here, we describe the methods for sample preparation, pretreatment, staining, clearing, and mounting for imaging.

Adipo: 三维脂肪组织成像的组织清除方法

Adipose tissue plays a central role in energy homeostasis and thermoregulation. It is composed of different types of adipocytes, as well as adipocyte ...

Reverse Dissection and DiceCT Reveal Otherwise Hidden Data in the Evolution of the Primate Face

Facial expressions, or facial displays, of social or emotional intent are produced by many mammalian taxa as a means of visually communicating with conspecifics at a close range. These displays are achieved by contraction of the mimetic muscles, which are skeletal muscle attached to the dermis of the face. Reverse dissection, removing the full facial mask from the skull and approaching mimetic muscles in reverse, is an effective but destructive way of revealing the morphology of mimetic muscles but it is destructive. DiceCT is a novel mechanism for visualizing skeletal muscles, including mimetic muscles, and isolating individual muscle fascicles for quantitative measurement. Additionally, DiceCT provides a non-destructive mechanism for visualizing muscles. The combined techniques of reverse dissection and DiceCT can be used to assess the evolutionary morphology of mimetic musculature as well as potential contraction strength and velocity in these muscles. This study further demonstrates that DiceCT can be used to accurately and reliably visualize mimetic muscles as well as reverse dissection and provide a non-destructive method for sampling mimetic muscles.

Facial expressions are a mode of visual communication produced by mimetic muscles. Here, we present protocols for the novel techniques of reverse dissection and DiceCT to fully visualize and assess mimetic muscles. These combined techniques can examine both morphological and physiological aspects of mimetic musculature to determine functional aspects.

灵长类动物脸演变过程中的反向解剖与 dicect 显示其他隐藏数据

Facial expressions, or facial displays, of social or emotional intent are produced by many mammalian taxa as a means of visually communicating with ...

A High Output Method to Isolate Cerebral Pericytes from Mouse

In recent years, cerebral pericytes have become the focus of extensive research in vascular biology and pathology. The importance of pericytes in blood brain barrier formation and physiology is now demonstrated but its molecular basis remains largely unknown. As the pathophysiological role of cerebral pericytes in neurological disorders is intriguing and of great importance, the in vitro models are not only sufficiently appropriate but also able to incorporate different techniques for these studies. Several methods have been proposed as in vitro models for the extraction of cerebral pericytes, although an antibiotic-free protocol with high output is desirable. Most importantly, a method that has increased output per extraction reduces the usage of more animals.
Here, we propose a simple and efficient method for extracting cerebral pericytes with sufficiently high output. The mouse brain tissue homogenate is mixed with a BSA-dextran solution for the separation of the tissue debris and microvascular pellet. We propose a three-step separation followed by filtration to obtain a microvessel rich filtrate. With this method, the quantity of microvascular fragments obtained from 10 mice is sufficient to seed 9 wells (9.6 cm2 each) of a 6-well plate. Most interestingly with this protocol, the user can obtain 27 pericyte rich wells (9.6 cm2 each) in passage 2. The purity of the pericyte cultures are confirmed with the expression of classical pericyte markers: NG2, PDGFR-&#946; and CD146. This method demonstrates an efficient and feasible in vitro tool for physiological and pathophysiological studies on pericytes.

We present a protocol for the extraction of murine cerebral pericytes. Based on an antibiotic-free enrichment oriented pericyte extraction, this protocol is a valuable tool for in vitro studies providing high purity and high yield, thus decreasing the number of experimental animals used.

一种从小鼠中分离大脑围取体的高输出方法

In recent years, cerebral pericytes have become the focus of extensive research in vascular biology and pathology. The importance of pericytes in blood ...

A Step-by-Step Guide to Mosquito Electroantennography

Female mosquitoes are the deadliest animals on earth, claiming the lives of more than 1 million people every year due to pathogens they transmit when acquiring a blood-meal. To locate a host to feed on, mosquitoes rely on a wide range of sensory cues, including visual, mechanical, thermal, and olfactory. The study details a technique, electroantennography (EAG), that allows researchers to assess whether the mosquitoes can detect individual chemicals and blends of chemicals in a concentration-dependent manner. When coupled with gas-chromatography (GC-EAG), this technique allows to expose the antennae to a full headspace/complex mixture and determines which chemicals present in the sample of interest, the mosquito can detect. This is applicable to host body odors as well as plant floral bouquets or other ecologically relevant odors (e.g., oviposition sites odorants). Here, we described a protocol that permits long durations of preparation responsiveness time and is applicable to both female and male mosquitoes from multiple genera, including Aedes, Culex, Anopheles, and Toxorhynchites mosquitoes. As olfaction plays a major part in mosquito-host interactions and mosquito biology in general, EAGs and GC-EAG can reveal compounds of interest for the development of new disease vector control strategies (e.g., baits). Complemented with behavioral assays, the valence (e.g., attractant, repellent) of each chemical can be determined.

The present article details a step-by-step protocol for successful and low noise electroantennograms in several genera of mosquitoes, including both females and males.

蚊子电图分步指南

Female mosquitoes are the deadliest animals on earth, claiming the lives of more than 1 million people every year due to pathogens they transmit when ...

Methods Article

到大脑的门户：剖析灵长类动物的眼睛