Name: immunofluorescent staining for visualization of heterochromatin associated proteins in drosophila salivary glands
Uploaded: 2021-08-21T14:30:00
Description: Watch this Scientific Journal Video about Immunofluorescent Staining for Visualization of Heterochromatin Associated Proteins in Drosophila Salivary Glands at JoVE.com

我们感谢马可·安东尼奥·罗萨莱斯·维加和阿贝尔·塞古拉拍摄了一些焦点图像，卡门·穆尼奥斯为媒体准备，阿图罗·皮门特尔博士，M.C安德烈斯·萨拉莱吉博士和来自LMNA的克里斯·伍德博士就显微镜的使用提供建议。

1. 第三星幼虫培养
<ol><li>加入100克酵母、100克未精制的全甘蔗糖、16克琼脂、10mL丙烯酸和14克明胶，准备1升标准介质。将除酵母以外的所有成分溶解在 800 mL 的自来水中，然后溶解酵母。高压灭菌立即30分钟。<ol>
<li>之后，让媒体冷却到60°C，并将丙酸添加到最终浓度0.01%。让瓶子站立，直到明胶形成。</li>
	</ol>
</li>
	<li>为了优化3o星内幼虫培养，首先收集5至10天的老年人，并将50（25男25女）放在?...

<ol>
	<li>Berger, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emil+Heitz,+a+true+epigenetics+pioneer.">Emil Heitz, a true epigenetics pioneer.</a> Nature Reviews Molecular Cell Biology. 20 (10), 572 (2019).</li><li>Irick, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+function+for+heterochromatin.">A new function for heterochromatin.</a> Chromosoma. 103 (1), 1-3 (1994).</li><li>Kasinathan, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Innovation+of+heterochromatin+functions+drives+rapid+evolution+of+essential+ZAD-ZNF+genes+in+Drosophila.">Innovation of heterochromatin functions drives rapid evolution of essential ZAD-ZNF genes in Drosophila.</a> Elife. , 1-31 (2020).</li><li>Lifschytz, E., Hareven, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterochromatin+markers:+Arrangement+of+obligatory+heterochromatin,+histone+genes+and+multisite+gene+families+in+the+interphase+nucleus+of+D.+melanogaster.">Heterochromatin markers: Arrangement of obligatory heterochromatin, histone genes and multisite gene families in the interphase nucleus of D. melanogaster.</a> Chromosoma. 86 (4), 443-455 (1982).</li><li>Eissenberg, J. C., Elgin, S. C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HP1a:+A+structural+chromosomal+protein+regulating+transcription.">HP1a: A structural chromosomal protein regulating transcription.</a> Trends in Genetics. 30 (3), 103-110 (2014).</li><li>Lee, Y. C. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pericentromeric+heterochromatin+is+hierarchically+organized+and+spatially+contacts+H3K9me2+islands+in+euchromatin.">Pericentromeric heterochromatin is hierarchically organized and spatially contacts H3K9me2 islands in euchromatin.</a> PLoS Genetics. 16 (3), 1-27 (2020).</li><li>Koryakov, D. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+SUUR+protein+is+involved+in+binding+of+SU+(VAR)3-9+and+methylation+of+H3K9+and+H3K27+in+chromosomes+of+Drosophila+melanogaster.">The SUUR protein is involved in binding of SU (VAR)3-9 and methylation of H3K9 and H3K27 in chromosomes of Drosophila melanogaster.</a> Chromosome Research. 19 (2), 235-249 (2011).</li><li>Cao, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+Histone+H3+Lysine+27+Methylation+in+Polycomb-Group+Silencing.">Role of Histone H3 Lysine 27 Methylation in Polycomb-Group Silencing.</a> Science. 298 (5595), 1039 (2002).</li><li>Hines, K. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Domains+of+heterochromatin+protein+1+required+for+Drosophila+melanogaster+heterochromatin+spreading.">Domains of heterochromatin protein 1 required for Drosophila melanogaster heterochromatin spreading.</a> Genetics. 182 (4), 967-977 (2009).</li><li>Elgin, S. C. R., Reuter, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Position-effect+variegation,+heterochromatin+formation,+and+gene+silencing+in+Drosophila.">Position-effect variegation, heterochromatin formation, and gene silencing in Drosophila.</a> Cold Spring Harbor Perspectives in Biology. 5 (8), 1-26 (2013).</li><li>Eissenberg, J. C., Elgin, S. C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HP1a:+A+structural+chromosomal+protein+regulating+transcription.">HP1a: A structural chromosomal protein regulating transcription.</a> Trends in Genetics. 30 (3), 103-110 (2014).</li><li>Meyer-Nava, S., Torres, A., Zurita, M., Valadez-graham, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+effects+of+dADD1+misexpression+in+chromatin+organization+and+transcription.">Molecular effects of dADD1 misexpression in chromatin organization and transcription.</a> BMC Molecular and Cell Biology. 2, 1-17 (2020).</li><li>Tennessen, J. M., Thummel, C. 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P., Bownes, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Staging+the+metamorphosis+of+Drosophila+melanogaster.">Staging the metamorphosis of Drosophila melanogaster.</a> Journal of Embryology and Experimental Morphology. 66 (1967), 57-80 (1981).</li><li>Larson, A. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liquid+droplet+formation+by+HP1&#945;+suggests+a+role+for+phase+separation+in+heterochromatin.">Liquid droplet formation by HP1&#945; suggests a role for phase separation in heterochromatin.</a> Nature. 547 (7662), 236-240 (2017).</li><li>Larson, A. G., Narlikar, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Role+of+Phase+Separation+in+Heterochromatin+Formation,+Function,+and+Regulation.">The Role of Phase Separation in Heterochromatin Formation, Function, and Regulation.</a> Biochemistry. 57 (17), 2540-2548 (2018).</li><li>Keenen, M. M., Larson, A. G., Narlikar, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualization+and+Quantitation+of+Phase-Separated+Droplet+Formation+by+Human+HP1&#945;.">Visualization and Quantitation of Phase-Separated Droplet Formation by Human HP1&#945;.</a> Methods in Enzymology. , (2018).</li><li>Lu, B. Y., Emtage, P. C., Duyf, B. J., Hilliker, a. J., Eissenberg, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterochromatin+protein+1+is+required+for+the+normal+expression+of+two+heterochromatin+genes+in+Drosophila.">Heterochromatin protein 1 is required for the normal expression of two heterochromatin genes in Drosophila.</a> Genetics. 155 (2), 699-708 (2000).</li><li>Marsano, R. M., Giordano, E., Messina, G., Dimitri, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+New+Portrait+of+Constitutive+Heterochromatin:+Lessons+from+Drosophila+melanogaster.">A New Portrait of Constitutive Heterochromatin: Lessons from Drosophila melanogaster.</a> Trends in Genetics. , (2019).</li><li>Strom, A. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phase+separation+drives+heterochromatin+domain+formation.">Phase separation drives heterochromatin domain formation.</a> Nature. 547 (7662), 241-245 (2017).</li><li>Sanulli, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HP1+reshapes+nucleosome+core+to+promote+phase+separation+of+heterochromatin.">HP1 reshapes nucleosome core to promote phase separation of heterochromatin.</a> Nature. 575 (7782), 390-394 (2019).</li><li>Dialynas, G., Delabaere, L., Chiolo, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arp2/3+and+Unc45+maintain+heterochromatin+stability+in+Drosophila+polytene+chromosomes.">Arp2/3 and Unc45 maintain heterochromatin stability in Drosophila polytene chromosomes.</a> Experimental Biology and Medicine. 244 (15), 1362-1371 (2019).</li><li>Kolesnikova, T. 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真核生物的细胞功能可以定义细胞核内的3D结构，由不同蛋白质与染色质和包括RNA在内的各种分子之间的相互作用得到支持。在过去三年中，具有相关性的生物凝结物，包括异色素，在确定相分离方面发挥了根本作用，促进了活跃和抑制性色度素16、17、18的独特核空间组织。 
异色素对于保持细胞功能和身?...

自早期研究Emil Heitz1以来，异色素一直被认为是细胞过程的重要调节器，如基因表达、染色体的美色和间质分离，以及维持基因组稳定性2、3、4。
异色素主要分为两种类型：具有特征定义重复序列的构成异色素，以及存在于特定染色体位点（如端粒和中位体）中的可转换元素。这种类型的异色素主要通过特定的平生标记，如赖氨酸9的地基或三甲基化的希石H3（H3K9me3）和异色素蛋白1a（HP1a）5，6的结合来表观遗传。另一方面，通过染色体的手臂，由发育沉默的基因7，8组成。代谢细胞中异色素块的免疫染色，或相间细胞中异色素聚合物的观察，在了解异色区形成和功能方面，揭示了许多光。
使用卓索菲拉作为模型系统，使得开发必要的工具，研究异色素，而无需使用电子显微镜10。由于位置效应变异的描述和异色素相关蛋白质（如HP1a）和组蛋白后转化改性蛋白的发现，许多小组已经开发出几种免疫造血技术，允许这些异色区域的可视化10，11。
这些技术基于特定抗体的使用，这些抗体能够识别异色素相关蛋白质或希石标记。对于每个细胞类型和抗体，必须经验确定固定和渗透条件。此外，如果使用额外的机械过程（如挤压技术），条件可能会有所不同。在这个协议中，我们描述了使用果蝇唾液腺来研究异色病。唾液腺具有多聚化细胞，包含超过1000份基因组，因此提供了大部分染色质特征的放大视图，除了卫星DNA和一些被复制的异色区域。然而，异色素区域很容易在多极染色体制剂中可视化，但挤压技术有时可能会破坏典型的染色质结合复合物或染色质结构。因此，整个唾液腺组织中蛋白质的免疫定位可以超越这些不良反应。我们使用这个协议来检测几个染色质结合蛋白，我们已经证明，这个协议结合突变的果蝇种群可用于研究异色素干扰12。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1.5 mL microcentrifuge tubes</td>
			<td>Axygen MCT-150-C</td>
			<td>11351904</td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>16% formaldehyde</td>
			<td>Thermo Scientific</td>
			<td>28908</td>
			<td></td>
		</tr>
		<tr>
			<td>AF1 Citifluor</td>
			<td>Ted pella</td>
			<td>19470</td>
			<td>25 mL</td>
		</tr>
		<tr>
			<td>BSA, Molecular Biology Grade</td>
			<td>Roche</td>
			<td>10735078001</td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>Complete, protease inhibitors Ultra EDTA-free 
			protease inhibitors</td>
			<td>Merck</td>
			<td>5892953001</td>
			<td></td>
		</tr>
		<tr>
			<td>Coverslip</td>
			<td>Corning</td>
			<td>CLS285022-200EA</td>
			<td>22x22, brand not critical</td>
		</tr>
		<tr>
			<td>DTT</td>
			<td>Sigma</td>
			<td>d9779</td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>Sigma</td>
			<td>E5134</td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>EGTA</td>
			<td></td>
			<td></td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>Glass slide</td>
			<td>Gold seal</td>
			<td>3011</td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>H3BO3</td>
			<td>Baker</td>
			<td>0084-01</td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>H3K9me3</td>
			<td>Abcam</td>
			<td>8889</td>
			<td></td>
		</tr>
		<tr>
			<td>HP1a</td>
			<td>Hybridoma Bank</td>
			<td>C1A9</td>
			<td>Product Form&#160;Concentrate 0.1 mL</td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Baker</td>
			<td>3040-01</td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Baker</td>
			<td>9070-03</td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sigma</td>
			<td>71376</td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>NaOH</td>
			<td></td>
			<td></td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>PIPES</td>
			<td></td>
			<td></td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>Rotator</td>
			<td>Thermo Scientific</td>
			<td>13-687-12Q</td>
			<td>&#160;Labquake Tube Shaker</td>
		</tr>
		<tr>
			<td>Thermo Mixer C</td>
			<td>Eppendorf</td>
			<td>13527550</td>
			<td>SmartBlock 1.5 mL</td>
		</tr>
		<tr>
			<td>Tris</td>
			<td>Milipore</td>
			<td>648311</td>
			<td>brand not critical</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma</td>
			<td>T8787</td>
			<td>100 mL, brand not critical</td>
		</tr>
		<tr>
			<td>&#946;-mercaptoethanol</td>
			<td>Bio-Rad</td>
			<td>1610710</td>
			<td>25 mL, brand not critical</td>
		</tr>
	</tbody>
</table>

immunofluorescent staining for visualization of heterochromatin associated proteins in drosophila salivary glands

通过免疫染色来可视化异色素聚合物可能具有挑战性。许多哺乳动物的染色质成分在 果蝇黑色素加斯特保存。 因此，研究异色素的形成和维护是一个很好的模型。 聚合细胞，如在第三星 D.黑色素幼 虫的唾液腺中发现的细胞，提供了观察血红素放大近千次的绝佳工具，并使研究人员能够研究异色素在细胞核中分布的变化。虽然异色素成分的观察可以直接在多极染色体制剂中进行，但某些蛋白质的定位可以通过治疗的严重程度来改变。因此，细胞中异色素的直接可视化补充了这种类型的研究。在此协议中，我们描述了用于此组织的免疫染色技术、使用二次荧光抗体和连续显微镜以更精确和更精细的方式观察这些异色素聚合物。

图1显示，在果蝇唾液腺中，HP1a免疫染色具有代表性。 一个积极的结果是观察一个焦点（图1a）（异色聚合物或凝结物）。负结果为无信号或分散信号。有时可以观察到双重信号，即双点（图1c），但它通常发生的数量较小。
数据分析可以表示为条形图，比较不同突变背景中 HP1a 的分布。例如，在 <stron...

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Immunofluorescent Staining for Visualization of Heterochromatin Associated Proteins in Drosophila Salivary Glands

此协议旨在可视化 多聚 体细胞中的异色素聚合物。

免疫荧光染色，用于 在果蝇 唾液腺中可视化异色素相关蛋白质

Visualization of heterochromatin aggregates by immunostaining can be challenging. Many mammalian components of chromatin are conserved in Drosophila ...

此协议的目的是可视化德罗索菲拉细胞中的异色素聚合物。为了达到这个目的，我们使用在第三星幼虫唾液腺中发现的多特尼细胞这些细胞的基因组被放大了近一千倍，染色体保存了界面细胞的特征。它们有定义明确的端粒，以及称为染色体的桥梁，其中所有的中心聚集，它主要是异色。使用第三个星内幼虫组织，也使我们能够评估不同遗传突变体背景中异色聚合的变化。为了优化第三个星内幼虫培养，您需要首先收集 5 到 10 天的老年人，并将大约 50 天放在标准媒体的宽颈瓶中。25男25女。将瓶子放在25摄氏度的受控温度孵化器中12小时。孵化期结束后，将成人取出并转移到新瓶中重复该程序。让胚胎在18摄氏度下生长22小时。对于幼虫收集，请确保您选择没有球形的流浪幼虫。解剖15对唾液腺，或在30分钟内在寒冷的PBS与显微镜下的抗议抑制剂尽可能多地。将唾液腺转移到另一个带冰冷 PBS 的管子。用冷 PBS 加抗议抑制剂洗一次。记住，总是等待组织到达管子的底部。从最后一步中取出 PBS 后，直接添加 0.5 毫升的 X 组牙龈固定缓冲和 50% 甲醇加 2% 甲醛。在摄氏四度下孵育两小时，轻度旋转。用特里斯-特里顿缓冲器进行一次五分钟的旋转洗涤，增加一毫升。始终等待组织到达管子底部。注意：等待组织到达管子底部。用特里斯-特里顿（Tris-Triton）孵育唾液腺，与上述相同，外加1%的β麦加普托乙醇，在37摄氏度下两小时，轻微摇晃。用BO3缓冲器清洗，在37摄氏度下用BO3加10毫摩尔DTT孵育，轻度摇晃15分钟。在潜伏期结束时，单独用一毫升 BO3 缓冲器进行洗涤。在室温下旋转两小时，将唾液腺在一毫升缓冲B中孵育两小时。删除所有缓冲 B 并添加缓冲 A 加感兴趣的抗体。在这种情况下，我们使用惠普一，A C 1 8 9 从Iridium银行。晚上三千分之一， 旋转四度。在这一点上，重要的是震动不会引发气泡，这可能会损害抗体。用缓冲B在室温下搅拌1毫米，用缓冲B洗涤15分钟。腺体与二级抗生素夫妇一起转移到缓冲B，在四度旋转下转诊两小时。在此步骤中，用铝纸箔覆盖管子以保护辅助抗体免受光线的照射非常重要。在室温下进行2x15分钟的垫圈，同时用一毫升缓冲器B.孵化，用DNA标记，如Sydox或OH，并在室温下溶解在一毫升缓冲B中10分钟，旋转。用缓冲 B 洗一次，用 PBS 洗一次，每次洗涤持续 10 分钟，同时在室温下旋转。最后，将唾液腺安装在滑梯上，使池面滑动。将唾液腺放在池中，并盖上西迪弗洛尔。为了避免气泡变形，将粘性液体扩展到所有地方。然后，用清晰的指甲油密封所有的滑梯。在荧光或聚焦显微镜下观察。如果当天不观察样品，则从灯光下储存四度。第一图显示了嗜血杆菌唾液腺中HB1单染的代表性结果。一个积极的结果是观察一个焦点，如在A，气动色聚合或冷凝水。负结果为无信号或分散信号。有时，双信号可以像 C 中的双点一样观察到，但通常以较小的数量发生。要分析数据，可以将其表示为条形图，比较具有不同突变背景的 HP1 的分布。例如，在第二图中，我们可以看到 98% 的核呈现一点分布。和 2% 的两个 foci 在野生类型。在突变体中，比例发生变化，两个 foci 的存在增加到 40%图三显示具有代表性的三甲基化赖氨酸 9H3 在单染色。导致果蝇唾液腺观察B中的一个焦点，这是一个有利的结果，在色聚合或凝结，在C的双重或三重信号可以看到在极少数情况下，但在少量。在本协议的末尾，您将能够看到HP1蛋白质的二位星凝结物，尽管状态的限制，这将是一个很好的第一种方法。

Research

JoVE Journal

Biology

Immunofluorescent Staining for Visualization of Heterochromatin Associated Proteins in Drosophila Salivary Glands

Dissection of Midgut and Salivary Glands from Ae. aegypti Mosquitoes

从AE肠和唾液腺的解剖。埃及伊蚊蚊

The mosquito midgut and salivary glands are key entry and exit points for pathogens such as Plasmodium parasites and Dengue viruses. This video protocol ...

科研

生物学

Staining Proteins in Gels

在凝胶染色蛋白质

Following separation by electrophoretic methods, proteins in a gel can be detected by several staining methods. This unit describes protocols for ...

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

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

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 ...

Purification of the M. magneticum Strain AMB-1 Magnetosome Associated Protein MamA&Delta;41

Purification of the M. magneticum Strain AMB-1 Magnetosome Associated Protein MamAÃƒÅ½Ã¢â‚¬Â41

纯化 M。 magneticum应变AMB - 1磁小体相关蛋白MamAΔ41

Magnetotactic bacteria comprise a diverse group of aquatic microorganisms that are able to orientate themselves along geomagnetic fields. This behavior is ...

Dissection and Staining of Drosophila Larval Ovaries

Dissection and Staining of Drosophila Larval Ovaries

夹层和染色果蝇幼虫卵巢

Many organs depend on stem cells for their development during embryogenesis and for maintenance or repair during adult life. Understanding how stem cells ...

Live Imaging of GFP-labeled Proteins in Drosophila Oocytes

Live Imaging of GFP-labeled Proteins in Drosophila Oocytes

GFP标记的蛋白质在实时成像<em&gt;果蝇</em&gt;卵母细胞

The  Drosophila oocyte has been established as a versatile system for investigating  fundamental questions such as cytoskeletal function, cell ...

In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

肌肉干细胞中自噬的原位免疫荧光染色

Increasing evidence points to autophagy as a crucial regulatory process to preserve tissue homeostasis. It is known that autophagy is involved in skeletal ...

Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins

This method provides a fast approach for the determination of plasma membrane partitioning of any fluorescently-tagged peripherally-associated protein ...

Visualization of 3D White Adipose Tissue Structure Using Whole-mount Staining

三维白色脂肪组织结构的全装染色可视化

Adipose tissue is an important metabolic organ with high plasticity and is responsive to environmental stimuli and nutrient status. As such, various ...

Dissection and Lipid Droplet Staining of Oenocytes in Drosophila Larvae

Dissection and Lipid Droplet Staining of Oenocytes in Drosophila Larvae

果蝇幼虫的欧诺细胞的解剖和脂滴染色

Lipids are essential for animal development and physiological homeostasis. Dysregulation of lipid metabolism results in various developmental defects and ...

免疫荧光染色，用于 在果蝇 唾液腺中可视化异色素相关蛋白质

免疫荧光染色，用于在果蝇唾液腺中可视化异色素相关蛋白质