<ol style="list-style-type:upper-alpha;"><li> 幻象病变的制备（约38至48小时，准备幻象FNA的能力（图1）实际执业会议之前或利用FNA的来源，美国准备使用“AV幻影”幻象病变（没有冷冻冷藏条件下储存）保持甚至3至4天前的准备练习。 </li><li>实践FNA的能力：虽然我们已经证明使用过程Shidham的THFV（收割机的组织与功能阀）（FNA的来源，美国），其他任何活检系统可用于练习（图2）。 </li><li>传播和处理的玻片上的吸（图3）。 </li></ol>所需材料<ol style="list-style-type:lower-alpha;"><li>清除硅橡胶填缝，可在很多施工/家装店。许多品牌都包括GETM，磷酸二铵，红魔鬼，白色的闪电，和当地的连锁品牌。我们用硅橡胶，白防雷产品，克利夫兰，美国俄亥俄州<a href="http://www.wlcaulk.com/products/all_purpose/silicone_rubber/wl09010.html">http://www.wlcaulk.com/products/all_purpose/silicone_rubber/wl09010.html</a> ） </li><li>压胶枪</li><li>一个一块保鲜膜板（我们使用萨兰保鲜加WRAP，美国庄臣父子公司，拉辛，威斯康星，美国<a href="http://www.saranbrands.com/plastic-wrap/index.asp">http://www.saranbrands.com/plastic-wrap/index.asp</a> </li><li>乳胶手套</li><li>宽直径的香蕉或香肠。 </li><li>事务所平底板（约6英寸x 6英寸）厚的卡板，亚克力板，或其他。 </li><li> FNA的表演设备<ul><li> 25计的注射针头</li><li> Shidham THFV（或执行FNA的其他组件） </li><li>载玻片</li><li>染色试剂盒</li></ul></li></ol>一步的步骤细节 A.准备（准备幻象1至2天前FNA的能力（图2）执业或利用FNA的来源，美国准备使用“AV幻影”幻影病变。 <ol><li>切6 × 6英寸大小支持板。 </li><li>剪下的“幻象病灶核心 ”（香蕉或香肠）的2.5英寸长的段。 </li><li>排出0.5英寸底板上的一层厚厚的填缝。 </li><li>轻轻按下“幻影病变核心”这层填缝。 </li><li>添加更多填缝“幻影病变核心”，因此，它是完全嵌入填缝土堆，没有捕获气泡。 </li><li>调整和形状的“幻象病灶核心 ”与戴手套的手的填缝。 </li><li>以保鲜膜一个6 × 6英寸大小的一块和覆盖填缝土堆，土堆表面用干净的，戴手套的手成光滑的凸配置调整。 </li><li>让填缝土堆治愈长于根据产品说明书中提到的固化时间为6-8小时，（通常为24小时），在温暖的地方。 </li><li>固化后，填缝应牢固，保鲜膜可以很容易揭下。 </li><li> FNA的幻象病变准备执业FNA程序后，约8小时时填缝整个土堆已经完全治愈的中心，包含“虚拟病变的核心 ”（香蕉或香肠） 。 </li></ol> B.实践AV幻影病变活检能力。 任何获得高品质的的方法，以优良的品质直接细胞学涂片足够的活检抽吸应解决以下关键点<ol style="list-style-type:upper-roman;"><li>针规，在程序的标题所暗示，更细的针头（高计数量）产生更好地吸出，以最小的血液稀释。关键平衡的优点和局限性，22日至25表针针最佳的机械稳定性，在不影响活检吸质量的首选。 </li><li>控制在适当的活检在真空中的应用和释放序列的过程，这是至关重要的，以便对针枢纽，推进每一针中风的采样细胞。然而，不应该被允许的吸气材料用注射器的针吸大会面积达到。如果这是无法阻止的，材料可能很难检索涂片（然而，这种材料可被用于细胞块制备或浓度的方法，如Cytospin，过滤器的方法，或最近的液基细胞学检查的准备工作）。 </li><li>发展适当的技能， 在吸气标本的处理，包括直接涂片制备及现场充足评估与辅助检查，如细 ​​胞块免疫细胞化学选修，选修淋巴组织增生性病变的免疫表型流式细胞仪准备适当分流，提交微生物的无菌通行证研究中，和队友（细胞遗传学等）的分子检测里亚尔。 </li></ol> 坝表演FNA程序（Shidham的THFV所示[1]（FNA的来源，美国）;然而，任何其他的FNA系统可用于练习）（图3,4,5） 。 <ol><li>阀门关闭，准备活检大会。 </li><li>找到病变部位，然后插入针尖在病灶。 </li><li>打开阀门，通过其尖端的针，连接真空吸代表病变的细胞成分 ，与每个到病变部位来回中风。 </li><li>针插入病变在不同的方向，保持对病灶的真空和样品的不同领域。 </li><li>关闭阀门，拔下针管腔内的真空。 </li><li>均衡的压力与环境条件（例如，从针头注射器阀一瞬间断开和重新连接，如果使用特殊装置内置的步骤，如“THFV&#39;，是，这和从BA1通过BA6其他步骤是一致的自动阀系统按照不同的步骤）[1]。 </li><li>从病灶取出针与阀关闭，以防止通过毛细作用进入病灶标本的损失，除非有轻微的负压力枢纽年底。 </li><li>打跑的载玻片上，吸气内容，准备直接涂片（见下文）（图6）。 </li></ol> BB。吸气标本适当分流处理 传播玻片吸和编制质量良好（图6）直接涂片，其次是适当的染色和分流（图7,8）。 <ol><li>一滴于载玻片上抛下吸传播的各种方法，如（图6）[2]。通常的做法是“A”。然而，如果试样是与一些microfragments稀释血液，多余的血是退掉倾斜幻灯片，然后microfragments类似&#39;A&#39;这个幻灯片和第二载玻片之间传播。如果吸主要是没有暂停，从淋巴组织增生性病变如microfragments，它是传播类似&#39;B&#39;或&#39;c&#39;。 </li><li>涂片可在各种不同的染色方法方法处理，并固定在现场充足评估。然而，最好的办法是风干所有的污迹。 Romanowski污渍（如Difff快客染色，瑞氏染色）后，甲醇固定，或与子宫颈抹片，风干涂片可染色，染色后生理盐水补液，后固定（在95％的乙醇或95％的乙醇最好用5 ％乙酸）[3]。由于模糊血消除干扰是一个明显的好处[3]。 </li><li>根据现场充足评估期间的初步cytomorphologic调查结果，建议额外的分流（如选修单元块，电子显微镜，微生物学文化，流式细胞仪，分子检测，细胞遗传学等）表示。 </li></ol>代表性的成果： <img border="0" src="/files/ftp_upload/1404/1404fig1tmb.jpg" alt="图1" /> <a href="https://www.jove.com/files/ftp_upload/1404/1404fig1.jpg">看到图1的放大版本，请点击</a>这里。 图1。准备影音幻影练FNA的能力所需的材料（*我们使用萨兰保鲜膜，没有坚持在年底填缝，而删除的嵌缝膏完全固化后。） <hr style="border-bottom:1px dotted #999999;" /><img border="0" src="/files/ftp_upload/1404/1404fig2tmb.jpg" alt="图2" /> <a href="https://www.jove.com/files/ftp_upload/1404/1404fig2.jpg">请点击这里看大图图2。</a> 图2准备影音幻影病变。 <hr style="border-bottom:1px dotted #999999;" /><img border="0" src="/files/ftp_upload/1404/1404fig3tmb.jpg" alt="图3" /> <a href="https://www.jove.com/files/ftp_upload/1404/1404fig3.jpg">图3看到一个更大的版本，请点击这里</a> 。 图3。活检过程中的关键步骤。 <hr style="border-bottom:1px dotted #999999;" /><img border="0" src="/files/ftp_upload/1404/1404fig4tmb.jpg" alt="图4" /> <a href="https://www.jove.com/files/ftp_upload/1404/1404fig4.jpg">图4看到一个更大的版本，请点击这里</a> 。 图4。EUS - FNA的关键步骤摘要*（见图3） <hr style="border-bottom:1px dotted #999999;" /><img border="0" src="/files/ftp_upload/1404/1404fig5tmb.jpg" alt="图5" /> <a href="https://www.jove.com/files/ftp_upload/1404/1404fig5.jpg">请点击这里看大图图5。</a> 图5。陷阱活检进程的关键步骤edure。 <hr style="border-bottom:1px dotted #999999;" /><img border="0" src="/files/ftp_upload/1404/1404fig6tmb.jpg" alt="图6" /> <a href="https://www.jove.com/files/ftp_upload/1404/1404fig6.jpg">请点击这里看大图图6。</a> 图6两个幻灯片之间的活检标本直接涂片的制备。 <hr style="border-bottom:1px dotted #999999;" /><img border="0" src="/files/ftp_upload/1404/1404fig7tmb.jpg" alt="图7" /> <a href="https://www.jove.com/files/ftp_upload/1404/1404fig7.jpg">图7看到一个更大的版本，请点击这里</a> 。 图7。湿固定与风干涂片。 <hr style="border-bottom:1px dotted #999999;" /><img border="0" src="/files/ftp_upload/1404/1404fig8tmb.jpg" alt="图8" /> <a href="https://www.jove.com/files/ftp_upload/1404/1404fig8.jpg">请点击这里看大图图8。</a> 图8。湿固定与风干涂片。

<ol>
	<li>Shidham, V. B., Pandit, A. W., Rao, R. N., Basir, Z., Shidham, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+Harvester+with+Functional+Valve+(THFV):+Shidham's+device+for+reproducibly+higher+specimen+yield+by+fine+needle+aspiration+biopsy+with+easy+to+perform+steps.">Tissue Harvester with Functional Valve (THFV): Shidham's device for reproducibly higher specimen yield by fine needle aspiration biopsy with easy to perform steps.</a> BMC Clinical Pathology. 7 (2), (2007).</li><li>Shidham, V. B., Epple, J., Shidham, V. B., Atkinson, B. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AppendiX+I:+Collection+and+processing+of+effusion+fluids+for+cytopathologic+evaluation.">AppendiX I: Collection and processing of effusion fluids for cytopathologic evaluation.</a> , 207-235 (2007).</li><li>Shidham, V., Kampalath, B., England, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Routine+air+drying+of+all+the+smears+prepared+during+fine+needle+aspiration+and+intraoperative+cytology+studies:+An+opportunity+to+practice+a+unified+protocol,+offering+the+flexibility+of+choosing+variety+of+staining+methods.">Routine air drying of all the smears prepared during fine needle aspiration and intraoperative cytology studies: An opportunity to practice a unified protocol, offering the flexibility of choosing variety of staining methods.</a> Acta Cytologica. (45), 60-68 (2001).</li></ol>

细针穿刺活检（FNAB）被广泛作为一种经济，安全，快速的微创组织各种肿瘤和病变的诊断技术。在此过程中，一个18至27号针头插入到病变部位，来回移动多次在病变部位的不同地区根据不同的强度，范围从最小的只有毛细作用形成约10毫升控制的真空（吸）真空注射器检索诊断组织材料。一个人数较多的细针头是首选检索良好的细胞学材料，准备直接涂片检查。更多针头18G检索更多的样本适合细胞块，流式细胞仪，电子显微镜，细胞遗传学，微生物学文化等，虽然过程很简单，控制各种适当的应用程序和真空释放在适当的步骤，协调一致地步骤是非常关键的为达到最佳效果。 由于专门设计的高收益针，传统的皮下注射器针头或修改的非可用性，是用来执行此过程。然而，注射针头有诸多限制，导致产量低的样品，如果用传统方式。特殊的注射器夹具的要求通常会增加程序的复杂性。因此，由于缺乏适合市售FNAB针设备，患有这种高度灵活的程序不足标本检索的一个经常出现的问题。近日，设备，提高产量与可重复性已经报道[1]。 此外，目前还缺乏一个合适的模型练习熟练卫生人员进行培训，包括居民和研究员FNAB。我们一块新鲜的牛肝脏乳胶手套塞进执业FNAB能力模型进行了标准化。然而，这种模式是很难在短时间内组织。采购新鲜的牛肝脏通常是限制的困境。此外，肝脏处理可能引进作为一项重大的借鉴污染的相关因素。乳胶手套的倾向，泄漏后几个针头刺破导致潜在的烂摊子，在练习。 我们的报告，并描述了一个新颖而简单的方法，市售的硅橡胶嵌缝膏和其他容易获得的材料中嵌入一个小块的香蕉（或香肠）。 FNAB幻象保留真空密封，从而使材料从嵌入式的标本，相当于病变在临床上的实际FNAB程序的愿望。在这段视频中，我们展示了FNAB幻象病变和实践FNAB程序的过程。这也表明在简短的检索标本的处理，包括涂片制备和染色[2,3]。 由于填缝需要时间来治愈，幻象病变应准备提前约24至48小时，执业会议之前。可比准备使用幻像病变可从商业来源（FNA的来源，美国）。 幻象病变执业FNAB能力，可用于培训的居民和研究员的患者在临床上表现FNAB开始前。

preparation and using phantom lesions to practice fine needle aspiration biopsies

目前，卫生工作者，包括居民和研究员没有一个合适的幻象模型，以实践的细针穿刺活检（FNAB）程序。在过去，我们的标准化模型乳胶手套含有新鲜牛肝脏执业FNAB组成。然而，这种模式是很难在短时间内与采购新鲜的牛肝脏中最具挑战性的的一个环节，组织和准备​​。肝脏处理与污染有关的问题，也是一个重要的拉回。此外，手套材料的泄漏通过几针后，造成的烂摊子。 我们已经建立了一个简单的方法，在市售的硅橡胶填缝嵌入一小块香肠或香蕉的小说。这种模式允许保留材料的真空密封和愿望嵌入式标本相似，临床上的肿块实际FNAB程序，。 针枢纽的吸气材料可以处理类似采购的标本，在实际FNAB过程中，促进涂片制备和染色额外的能力。

Watch this Scientific Journal Video about Preparation and Using Phantom Lesions to Practice Fine Needle Aspiration Biopsies at JoVE.com

Preparation and Using Phantom Lesions to Practice Fine Needle Aspiration Biopsies

细针穿刺活检（FNAB）执业学员是比较具有挑战性的，由于缺乏一个容易获得的，适当的病变。准备的AV幻象执业FNAB程序和掌握的熟练程度的病变是比较容易的。

制备及使用幻影病变实践细针穿刺活检

Currently, health workers including residents and fellows do not have a suitable phantom model to practice the fine- needle aspiration biopsy (FNAB) ...

preparation-using-phantom-lesions-to-practice-fine-needle-aspiration

Research

JoVE Journal

Biology

17.7K 次观看.  University of Wisconsin - Milwaukee. 细针穿刺活检（FNAB）执业学员是比较具有挑战性的，由于缺乏一个容易获得的，适当的病变。准备的AV幻象执业FNAB程序和掌握的熟练程度的病变是比较容易的。

视频: Preparation and Using Phantom Lesions to Practice Fine Needle Aspiration Biopsies

Gross and Fine Dissection of Inner Ear Sensory Epithelia in Adult Zebrafish (Danio rerio)

Neurosensory epithelia in the inner ear are the crucial structures for hearing and balance functions. Therefore, it is important to understand the cellular and molecular features of the epithelia, which are mainly composed of two types of cells: hair cells (HCs) and supporting cells (SCs). Here we choose to study the inner ear sensory epithelia in adult zebrafish not only because the epithelial structures are highly conserved in all vertebrates studied, but also because the adult zebrafish is able to regenerate HCs, an ability that mammals lose shortly after birth. We use the inner ear of adult zebrafish as a model system to study the mechanisms of inner ear HC regeneration in adult vertebrates that could be helpful for clinical therapy of hearing/balance deficits in human as a result of HC loss.
Here we demonstrate how to do gross and fine dissections of inner ear sensory epithelia in adult zebrafish. The gross dissection removes the tissues surrounding the inner ear and is helpful for preparing tissue sections, which allows us to examine the detailed structure of the sensory epithelia. The fine dissection cleans up the non-sensory-epithelial tissues of each individual epithelium and enables us to examine the heterogeneity of the whole epithelium easily in whole-mount epithelial samples.

Gross and Fine Dissection of Inner Ear Sensory Epithelia in Adult Zebrafish Danio rerio

The inner ear sensory epithelium of adult zebrafish is a good model system for understanding the mechanisms of hair cell regeneration in adult vertebrates. This protocol demonstrates the fine dissection of the epithelia, through which we can get tissue samples for studying the regenerative events at cellular and subcellular levels.

格罗斯和精细解剖内耳感觉上皮细胞在成年斑马鱼（斑马鱼）

Neurosensory epithelia in the inner ear are the crucial structures for hearing and balance functions. Therefore, it is important to understand the ...

科研

生物学

Western Blotting: Sample Preparation to Detection

Western blotting is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide (denaturing conditions) or by the 3-D structure of the protein (native/ non-denaturing conditions). The proteins are then transferred to a membrane (typically nitrocellulose or PVDF), where they are probed (detected) using antibodies specific to the target protein.

Western blotting is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract.

免疫印迹检测样品制备

Western blotting is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract. It uses gel ...

Advanced Confocal Microscopy Techniques to Study Protein-protein Interactions and Kinetics at DNA Lesions

Local microirradiation with lasers represents a useful tool for studies of DNA-repair-related processes in live cells. Here, we describe a methodological approach to analyzing protein kinetics at DNA lesions over time or protein-protein interactions on locally microirradiated chromatin. We also show how to recognize individual phases of the cell cycle using the Fucci cellular system to study cell-cycle-dependent protein kinetics at DNA lesions. A methodological description of the use of two UV lasers (355 nm and 405 nm) to induce different types of DNA damage is also presented. Only the cells microirradiated by the 405-nm diode laser proceeded through mitosis normally and were devoid of cyclobutane pyrimidine dimers (CPDs). We also show how microirradiated cells can be fixed at a given time point to perform immunodetection of the endogenous proteins of interest. For the DNA repair studies, we additionally describe the use of biophysical methods including FRAP (Fluorescence Recovery After Photobleaching) and FLIM (Fluorescence Lifetime Imaging Microscopy) in cells with spontaneously occurring DNA damage foci. We also show an application of FLIM-FRET (Fluorescence Resonance Energy Transfer) in experimental studies of protein-protein interactions.

Laser microirradiation is a useful tool for studies of DNA repair in living cells. A methodological approach for the use of UVA lasers to induce various DNA lesions is shown. We have optimized a method for local microirradiation that maintains the normal cell cycle; thus, irradiated cells proceed through mitosis.

高级共焦显微镜技术研究 DNA 损伤的蛋白质-蛋白质相互作用和动力学

Local microirradiation with lasers represents a useful tool for studies of DNA-repair-related processes in live cells. Here, we describe a methodological ...

Immunolabelling Myofiber Degeneration in Muscle Biopsies

The necrosis of muscle fibres (myonecrosis) plays a central role in the pathogenesis of several muscle conditions, including muscular dystrophies. Therapeutic options addressing the causes of muscular dystrophy pathogenesis are expected to alleviate muscle degeneration. Therefore, a method to assay and quantify the extent of cell death in muscle biopsies is needed. Conventional methods to observe myofiber degeneration in situ are either poorly quantitative or rely on the injection of vital dyes. In this article, an immunofluorescence protocol is described that stains necrotic myofibers by targeting immunoglobulin G (IgG) uptake by myofibers. The IgG uptake method is based on cell features characterizing the necrotic demise, including 1) the loss of plasma membrane integrity with the release of damage-associated molecular patterns and 2) the uptake of plasmatic proteins. In murine cross-sections, the co-immunolabelling of myofibers, extracellular matrix proteins, and mouse IgG allows clean and straightforward identification of myofibers with necrotic fate. This simple method is suitable for quantitative analysis and applicable to all species, including human samples, and does not require the injection of vital dye. The staining of necrotic myofibers by IgG uptake can also be paired with other co-immunolabelling.

Described here is a protocol for direct immunolabelling of necrotic myofibers in muscle cryosections. Necrotic cells are permeable to serum proteins, including immunoglobulin G (IgG). Revealing the uptake of IgG by myofibers allows the identification and quantification of myofibers that undergo necrosis regardless of muscle condition.

肌肉活检中的免疫标记肌纤维退化

The necrosis of muscle fibres (myonecrosis) plays a central role in the pathogenesis of several muscle conditions, including muscular dystrophies. ...

Single Cell Micro-aspiration as an Alternative Strategy to Fluorescence-activated Cell Sorting for Giant Virus Mixture Separation

During the amoeba co-culture process, more than one virus may be isolated in a single well. We previously solved this issue by end point dilution and/or fluorescence activated cell sorting (FACS) applied to the viral population. However, when the viruses in the mixture have similar morphologic properties and one of the viruses multiplies slowly, the presence of two viruses is discovered at the stage of genome assembly and the viruses cannot be separated for further characterization. To solve this problem, we developed a single cell micro-aspiration procedure that allows for separation and cloning of highly similar viruses. In the present work, we present how this alternative strategy allowed us to separate the small viral subpopulations of Clandestinovirus ST1 and Usurpativirus LCD7, giant viruses that grow slowly and do not lead to amoebal lysis compared to the lytic and fast-growing Faustovirus. Purity control was assessed by specific gene amplification and viruses were produced for further characterization.

Here, we describe a single cell micro-aspiration method for the separation of infected amoebae. In order to separate viral subpopulations in Vermamoeba vermiformis infected by Faustoviruses and unknown giant viruses, we developed the protocol detailed below and demonstrated its ability to separate two low-abundance novel giant viruses.

单细胞微吸气作为荧光激活细胞分类的替代策略，用于巨型病毒混合物分离

During the amoeba co-culture process, more than one virus may be isolated in a single well. We previously solved this issue by end point dilution and/or ...

Assessing Agrochemical Risk to Mated Honey Bee Queens

Current risk assessment strategies for honey bees rely heavily upon laboratory tests performed on adult or immature worker bees, but these methods may not accurately capture the effects of agrochemical exposure on honey bee queens. As the sole producer of fertilized eggs inside a honeybee colony, the queen is arguably the most important single member of a functioning colony unit. Therefore, understanding how agrochemicals affect queen health and productivity should be considered a critical aspect of pesticide risk assessment. Here, an adapted method is presented to expose honey bee queens and worker queen attendants to agrochemical stressors administered through a worker diet, followed by tracking egg production in the laboratory and assessing first instar eclosion using a specialized cage, referred to as a Queen Monitoring Cage. To illustrate the method&#39;s intended use, results of an experiment in which worker queen attendants were fed diet containing sublethal doses of imidacloprid and effects on queens were monitored are described.

This protocol was developed to enhance the understanding of how agrochemicals affect honey bee (Apis mellifera) reproduction by establishing methods to expose honey bee queens and their worker caretakers to agrochemicals in a controlled, laboratory setting and carefully monitoring their relevant responses.

评估交配蜜蜂女王的农用化学品风险

Current risk assessment strategies for honey bees rely heavily upon laboratory tests performed on adult or immature worker bees, but these methods may not ...

Profiling of H3K4me3 Modification in Plants using Cleavage under Targets and Tagmentation

Epigenomic regulation at the chromatic level, including DNA and histone modifications, behaviors of transcription factors, and non-coding RNAs with their recruited proteins, lead to temporal and spatial control of gene expression. Cleavage under targets and tagmentation (CUT&#38;Tag) is an enzyme-tethering method in which the specific chromatin protein is firstly recognized by its specific antibody, and then the antibody tethers a protein A-transposase (pA-Tn5) fusion protein, which cleaves the targeted chromatin in situ by the activation of magnesium ions. Here, we provide our previously published CUT&#38;Tag protocol using intact nuclei isolated from allortetraploid cotton leaves with modification. This step-by-step protocol can be used for epigenomic research in plants. In addition, substantial modifications for plant nuclei isolation are provided with critical comments.

Cleavage under targets and tagmentation (CUT&amp;Tag) is an efficient chromatin epigenomic profiling strategy. This protocol presents a refined CUT&amp;Tag strategy for the profiling of histone modifications in plants.

在靶标和标记下使用裂解分析植物中的H3K4me3修饰

Epigenomic regulation at the chromatic level, including DNA and histone modifications, behaviors of transcription factors, and non-coding RNAs with their ...

An Improved Chemotaxis Assay for the Rapid Identification of Rhizobacterial Chemoattractants in Root Exudates

Chemotaxis identification is very important for the research and application of rhizosphere growth-promoting bacteria. We established a straightforward method to quickly identify the chemoattractants that could induce the chemotactic movement of rhizosphere growth-promoting bacteria on sterile glass slides via simple steps. Bacteria solution (OD600 = 0.5) and sterile chemoattractant aqueous solution were added dropwise on the glass slide at an interval of 1 cm. An inoculating loop was used to connect the chemoattractant aqueous solution to the bacterial solution. The slide was kept at room temperature for 20 min on the clean bench. Finally, the chemoattractant aqueous solution was collected for bacterial counting and microscopic observation. In this study, through multiple comparisons of experimental results, the method overcame multiple shortcomings of traditional bacterial chemotaxis methods. The method reduced the error of plate counting and shortened the experimental cycle. For the identification of chemoattractant substances, this new method can save 2-3 days compared to the traditional method. Additionally, this method allows any researcher to systematically complete a bacterial chemotaxis experiment within 1-2 days. The protocol can be considered a valuable resource for understanding plant-microbe interactions.

Here, we present an improved chemotaxis assay protocol. The goal of this protocol is to reduce the steps and costs of traditional bacterial chemotaxis methods and to serve as a valuable resource for understanding plant-microbe interactions.

一种用于快速鉴定根系分泌物中根细菌化学引诱剂的改进趋化性测定

Chemotaxis identification is very important for the research and application of rhizosphere growth-promoting bacteria. We established a straightforward ...

Comprehensive Echocardiographic Assessment of Right Ventricle Function in a Rat Model of Pulmonary Arterial Hypertension

Pulmonary arterial hypertension (PAH) is a progressive disease caused by vasoconstriction and remodeling of the small arteries in the lungs. This remodeling leads to increased pulmonary vascular resistance, worsened right ventricular function, and premature death. Currently approved therapies for PAH largely target pulmonary vasodilator pathways; however, recent emerging therapeutic modalities are focused on other novel pathways involved in the pathogenesis of the disease, including right ventricle (RV) remodeling. Imaging techniques that allow longitudinal assessment of novel therapeutics are very useful for determining the efficacy of new drugs in preclinical studies. Noninvasive trans-thoracic echocardiography remains the standard approach to evaluating heart function and is widely used in rodent models. However, echocardiographic evaluation of the RV can be challenging due to its anatomical position and structure. In addition, standardized guidelines are lacking for echocardiography&#160;in preclinical rodent models, making it difficult to carry out a uniform assessment of RV&#160;function across studies in different laboratories. In preclinical studies, the monocrotaline (MCT) injury model in rats is widely used to evaluate drug efficacy for treating PAH. This protocol describes the echocardiographic evaluation of the RV in na&#239;ve and MCT-induced PAH rats.

The present protocol describes the echocardiographic characterization of right ventricular morphology and function in a rat model of pulmonary arterial hypertension.

肺动脉高压大鼠模型中右心室功能的综合超声心动图评估

Pulmonary arterial hypertension (PAH) is a progressive disease caused by vasoconstriction and remodeling of the small arteries in the lungs. This ...

使用皮肤活检生成患者来源的足细胞

Generation of Patient-Derived Podocytes from Skin Biopsies

Podocytes are epithelial cells sitting on the urinary site of the glomerular filtration barrier that contribute to the selective filter function of the glomerulus. Mutations in podocyte-specific genes can cause focal segmental glomerulosclerosis (FSGS), and podocytes are also affected in many other primary and secondary nephropathies. Due to their differentiated nature, primary cell culture models are limited for podocytes.&#160;Therefore, commonly conditionally immortalized cells are used. However, these conditionally immortalized podocytes (ciPodocytes) have several limitations: the cells can dedifferentiate in culture, especially when they reach confluency, and several podocyte-specific markers are either only slightly or not expressed at all. This brings the use of ciPodocytes and their applicability for physiological, pathophysiological, and clinical reach into question. Here, we describe a protocol for the generation of human podocytes-including patient-specific podocytes-from a skin punch biopsy by episomal reprogramming of dermal fibroblasts into hiPSCs and subsequent differentiation into podocytes. These podocytes resemble in vivo podocytes much better in terms of morphological characteristics, like the development of foot processes and the expression of the podocyte-specific marker. Finally, yet importantly, these cells maintain patients&#39; mutations, resulting in an improved ex vivo model to study podocyte diseases and potential therapeutic substances in an individualized approach.

作者聚焦：从皮肤活检中生成患者来源的足细胞  

This manuscript describes a two-step protocol to generate patient-specific podocytes from dermal fibroblasts via episomal reprogramming into human-induced pluripotent stem cells (hiPSCs) and subsequent differentiation into podocytes.

从皮肤活检中生成患者来源的足细胞

Podocytes are epithelial cells sitting on the urinary site of the glomerular filtration barrier that contribute to the selective filter function of the ...