Name: microwaving and fluorophore-tyramide for multiplex immunostaining on mouse adrenals &#8722; using unconjugated primary antibodies from the same host species
Uploaded: 2020-02-21T14:00:00
Description: Watch this Scientific Journal Video about Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals Ã¢Ë†' Using Unconjugated Primary Antibodies from the Same Host Species at

这项工作由NIH R00 HD032636支持。

1. 使用第一抗体染色
<ol><li>脱蜡和再水化 FFPE 滑动，每个步骤分配 5 分钟：二甲苯或等效试剂 3x、100% 乙醇 2x、95% 乙醇 1x、70% 乙醇 1x、50% 乙醇 1x 和蒸馏水 2x。 注：滑轨应从此补液步骤开始保持湿润，直到安装到最后一步。</li>
	<li>对于可选的抗原检索，将幻灯片放入 275 mL 的沸腾柠子钠溶液（10 mM，pH = 6.0）8分钟。为了保持溶液沸腾，将幻灯片平放在 14 x 9.5 x 9 cm3 （W x L x H） 移液器?...

<ol>
	<li>Lewis Carl, S. A., Gillete-Ferguson, I., Ferguson, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+indirect+immunofluorescence+procedure+for+staining+the+same+cryosection+with+two+mouse+monoclonal+primary+antibodies.">An indirect immunofluorescence procedure for staining the same cryosection with two mouse monoclonal primary antibodies.</a> Journal of Histochemistry and Cytochemistry. 41 (8), 1273-1278 (1993).</li><li>Lan, H. Y., Mu, W., Nikolic-Paterson, D. J., Atkins, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel,+simple,+reliable,+and+sensitive+method+for+multiple+immunoenzyme+staining:+use+of+microwave+oven+heating+to+block+antibody+crossreactivity+and+retrieve+antigens.">A novel, simple, reliable, and sensitive method for multiple immunoenzyme staining: use of microwave oven heating to block antibody crossreactivity and retrieve antigens.</a> Journal of Histochemistry and Cytochemistry. 43 (1), 97-102 (1995).</li><li>Tornehave, D., Hougaard, D. M., Larsson, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microwaving+for+double+indirect+immunofluorescence+with+primary+antibodies+from+the+same+species+and+for+staining+of+mouse+tissues+with+mouse+monoclonal+antibodies.">Microwaving for double indirect immunofluorescence with primary antibodies from the same species and for staining of mouse tissues with mouse monoclonal antibodies.</a> Histochemistry and Cell Biology. 113 (1), 19-23 (2000).</li><li>Kim, M., Soontornniyomkij, V., Ji, B., Zhou, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=System-wide+immunohistochemical+analysis+of+protein+co-localization.">System-wide immunohistochemical analysis of protein co-localization.</a> PLoS One. 7 (2), e32043 (2012).</li><li>Bolognesi, M. M., 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=Multiplex+Staining+by+Sequential+Immunostaining+and+Antibody+Removal+on+Routine+Tissue+Sections.">Multiplex Staining by Sequential Immunostaining and Antibody Removal on Routine Tissue Sections.</a> Journal of Histochemistry and Cytochemistry. 65 (8), 431-444 (2017).</li><li>Toth, Z. E., Mezey, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+visualization+of+multiple+antigens+with+tyramide+signal+amplification+using+antibodies+from+the+same+species.">Simultaneous visualization of multiple antigens with tyramide signal amplification using antibodies from the same species.</a> Journal of Histochemistry and Cytochemistry. 55 (6), 545-554 (2007).</li><li>Buchwalow, I., Samoilova, V., Boecker, W., Tiemann, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+immunolabeling+with+antibodies+from+the+same+host+species+in+combination+with+tyramide+signal+amplification.">Multiple immunolabeling with antibodies from the same host species in combination with tyramide signal amplification.</a> Acta Histochemica. 120 (5), 405-411 (2018).</li><li>Bauer, M., Schilling, N., Spanel-Borowski, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limitation+of+microwave+treatment+for+double+immunolabelling+with+antibodies+of+the+same+species+and+isotype.">Limitation of microwave treatment for double immunolabelling with antibodies of the same species and isotype.</a> Histochemistry and Cell Biology. 116 (3), 227-232 (2001).</li><li>Pirici, D., 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=Antibody+elution+method+for+multiple+immunohistochemistry+on+primary+antibodies+raised+in+the+same+species+and+of+the+same+subtype.">Antibody elution method for multiple immunohistochemistry on primary antibodies raised in the same species and of the same subtype.</a> Journal of Histochemistry and Cytochemistry. 57 (6), 567-575 (2009).</li><li>Glass, G., Papin, J. A., Mandell, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SIMPLE:+a+sequential+immunoperoxidase+labeling+and+erasing+method.">SIMPLE: a sequential immunoperoxidase labeling and erasing method.</a> Journal of Histochemistry and Cytochemistry. 57 (10), 899-905 (2009).</li><li>Zhang, W., 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=Fully+automated+5-plex+fluorescent+immunohistochemistry+with+tyramide+signal+amplification+and+same+species+antibodies.">Fully automated 5-plex fluorescent immunohistochemistry with tyramide signal amplification and same species antibodies.</a> Laboratory Investigation. 97 (7), 873-885 (2017).</li><li>Paul, A., Laufer, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endogenous+biotin+as+a+marker+of+adrenocortical+cells+with+steroidogenic+potential.">Endogenous biotin as a marker of adrenocortical cells with steroidogenic potential.</a> Molecular and Cellular Endocrinology. 336 (1-2), 133-140 (2011).</li><li>Bussolati, G., Gugliotta, P., Volante, M., Pace, M., Papotti, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retrieved+endogenous+biotin:+a+novel+marker+and+a+potential+pitfall+in+diagnostic+immunohistochemistry.">Retrieved endogenous biotin: a novel marker and a potential pitfall in diagnostic immunohistochemistry.</a> Histopathology. 31 (5), 400-407 (1997).</li><li>Wang, H., Pevsner, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+endogenous+biotin+in+various+tissues:+novel+functions+in+the+hippocampus+and+implications+for+its+use+in+avidin-biotin+technology.">Detection of endogenous biotin in various tissues: novel functions in the hippocampus and implications for its use in avidin-biotin technology.</a> Cell and Tissue Research. 296 (3), 511-516 (1999).</li></ol>

多路免疫染色可用于检查两种或两种以上抗原的细胞共定位。当初级抗体与不同的检测器（直接染色）结合时，这种广泛使用的技术提供了令人信服的共定位结果。然而，与间接染色相比，直接染色通常提供较弱的信号，这涉及结合的二级抗体来检测初级抗体。在间接染色中，高质量的多路免疫染色结果取决于二级抗体能否区分不同的原抗体。由于可能的抗体交叉反应，使用间接染色方法的多重?...

在多路免疫染色中，使用结合原抗体直接染色可提供信息性结果。不使用二级抗体，直接染色方法具有来自抗体交叉反应的虚假共定位信号的低风险。然而，在原抗体上的偶联记者（荧光、酶）或生物锡限制了其未来用途。或者，间接免疫染色通常通过使用未结合的初级抗体与标记的次级抗体提供更强的信号。理想情况下，用于多倍免疫染色的未结合原抗体应来自不同宿主物种，以避免抗体交叉反应。然而，不同宿主物种的初级抗体的适当组合并不总是可用的。
已经建立了几种方法来消除二级抗体与不需要的初级抗体反应的风险。一种常见的方法是使用F（ab）单体抗体在第二原抗体1染色之前，阻断第一原抗体复合物上任何剩余的结合表位。抗体剥离，这是类似于条带和重新探针的西方斑点片，去除以前染色的抗体复合物，而不剥离可检测的检测报告分子的沉积，如3，3'-二氨基苯甲二甲酸酯四氯化物（DAB）2和荧光酪酰胺沉积3。使用这种方法，不同颜色的记者从分子可以在同一张幻灯片上显示多路复用结果。通过完全去除先前沉积的抗体层，以及从其他抗体4、5中对齐随后获得的图像，也可以实现多重染色。这些方法都提供了可靠的结果，尽管每种方法都有其局限性，并且需要复杂的过程或特殊的成像系统。
本协议显示了使用常用缓冲液的抗体剥离方法的应用。该协议可用于对正式固定石蜡嵌入 （FFPE） 小鼠肾上腺部分执行多倍免疫荧光染色，同时使用来自同一宿主物种的两种未标记的主要抗体。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Antifade Mounting Medium</td>
			<td>Vector Laboratories</td>
			<td>H-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Biotinylated donkey anti-mouse</td>
			<td>JacksonImmuno</td>
			<td>715-066-151</td>
			<td>1:500 dilution</td>
		</tr>
		<tr>
			<td>Biotinylated donkey anti-rabbit</td>
			<td>JacksonImmuno</td>
			<td>711-066-152</td>
			<td>1:500 dilution</td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>BioLegend</td>
			<td>422801</td>
			<td>2 &#956;g/mL in distilled water</td>
		</tr>
		<tr>
			<td>Fluorescence microscope</td>
			<td>ECHO</td>
			<td>Revolve 4</td>
			<td></td>
		</tr>
		<tr>
			<td>Horseradish peroxidase-conjugated streptavidin</td>
			<td>JacksonImmuno</td>
			<td>016-030-084</td>
			<td>1:1000 dilution</td>
		</tr>
		<tr>
			<td>Microwave oven, 700W</td>
			<td>General Electric</td>
			<td>JEM3072DH1BB</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse anti-CYP2F2</td>
			<td>Santa Cruz,</td>
			<td>SC-374540</td>
			<td>1:250 dilution</td>
		</tr>
		<tr>
			<td>Mouse anti-TH</td>
			<td>Santa Cruz,</td>
			<td>SC-25269</td>
			<td>1:1000 dilution</td>
		</tr>
		<tr>
			<td>Normal donkey serum</td>
			<td>JacksonImmuno</td>
			<td>017-000-121</td>
			<td>2% serum in PBST</td>
		</tr>
		<tr>
			<td>Rabbit anti-20&#945;HSD</td>
			<td>Kerafast,</td>
			<td>EB4002</td>
			<td>1:500 dilution</td>
		</tr>
		<tr>
			<td>Rabbit anti-3&#946;HSD</td>
			<td>TransGenic,</td>
			<td>KO607</td>
			<td>1:250 dilution</td>
		</tr>
		<tr>
			<td>Rabbit anti-TH</td>
			<td>NOVUS,</td>
			<td>NB300-109</td>
			<td>1:1000 dilution</td>
		</tr>
		<tr>
			<td>Rabbit anti-&#946;-catenin</td>
			<td>Abcam,</td>
			<td>ab32572</td>
			<td>1:500 dilution</td>
		</tr>
		<tr>
			<td>Streptavidin Horseradish Peroxidase (SA-HRP)</td>
			<td>JacksonImmuno</td>
			<td>016-303-084</td>
			<td>1:1000 dilution</td>
		</tr>
		<tr>
			<td>TSA Cy3 Tyramide</td>
			<td>PerkinElmer</td>
			<td>SAT704B001EA</td>
			<td>1:100 dilution</td>
		</tr>
		<tr>
			<td>TSA Fluorescein Tyramide</td>
			<td>PerkinElmer</td>
			<td>SAT701001EA</td>
			<td>1:100 dilution</td>
		</tr>
	</tbody>
</table>

microwaving and fluorophore-tyramide for multiplex immunostaining on mouse adrenals &#8722; using unconjugated primary antibodies from the same host species

免疫染色在生物医学研究中广泛应用，以显示特定蛋白质的细胞表达模式。多路免疫染色允许使用多种原抗体进行标记。为了尽量减少抗体交叉反应，使用间接染色进行多路免疫染色需要来自不同宿主物种的未标记原抗体。然而，不同物种抗体的适当组合并不总是可用的。在这里，我们描述了一种使用来自同一宿主物种（例如，在本例中两种抗体来自兔子）的未标记原抗体的方法，用于在形式固定的石蜡嵌入（FFPE）小鼠肾上腺部分进行多倍免疫荧光。该方法使用抗原检索步骤中使用的相同程序和试剂来去除先前染色的原抗体复合物的活性。幻灯片使用一般免疫染色方案染色的第一个初级抗体，然后与生物浸化二级抗体结合步骤。然后，采用一种以氟磷-酪酰胺为基质的avidin-生物素-过氧化物酶信号开发方法。通过浸入微波沸腾柠酸钠溶液8分钟，剥离了第一原发抗体复合物的免疫活性。不溶性氟磷-酪酰胺沉积留在样品上，使滑道上沾染了其他主要抗体。虽然这种方法消除了大多数误报信号，但抗体交叉反应的某些背景可能仍然存在。如果样品富含内源性生物锡，则可以使用过氧化物酶结合二级抗体替代生物异化二级抗体，以避免从回收的内源性生物锡中误化的假阳性。

结果来自经过所有步骤处理的样本，包括抗原检索步骤1.2。这里使用的所有二级抗体都是生物异位。氟磷-酪酰胺用于从第一和第二原抗体产生信号。使用配备 FITC 立方体（用于绿色荧光）、TxRED 立方体（用于 Cy3）和 DAPI 立方体（用于 DAPI）的荧光显微镜捕获图像。
微摆动介导剥离消除了交叉反应性。 小鼠FFPE肾...

Watch this Scientific Journal Video about Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals Ã¢Ë†' Using Unconjugated Primary Antibodies from the Same Host Species at 

Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals ÃƒÂ¢Ã‹â€ ' Using Unconjugated Primary Antibodies from the Same Host Species

利用来自同一宿主物种的初级抗体，已经建立了几种不同的多路免疫染色方法。在这里，我们描述了在正式固定石蜡嵌入小鼠肾上腺部分进行多倍免疫染色期间，使用微波介导的抗体剥离和荧光素-酪酰胺来阻止抗体的交叉反应。

微挥和荧光素-提拉美化物，用于小鼠肾上腺的多重免疫染色 – 使用来自同一宿主物种的未结合原抗体

Immunostaining is widely used in biomedical research to show the cellular expression pattern of a given protein. Multiplex immunostaining allows labeling ...

免疫污染在生物医学研究中被广泛用于识别感兴趣的蛋白质的位置。多路复用免疫处理可以使用不同的原抗体检测同一样品上的多个靶点。这种方法的好处是使用常用的缓冲液来消除抗体交叉反应，允许使用来自同一宿主物种的两种或两种（多个未标记的原抗体）进行免疫控制。展示这个程序的将是研究生郑索菲娅和我实验室的客座学者琼霞。在染色之前，去蜡和补水形式固定石蜡嵌入幻灯片与五分钟的浸入每个解决方案，如指示。第二次蒸馏水浸泡后，将幻灯片放入275毫升沸腾的柠檬酸钠溶液中，放在带盖子的移液器尖端盒底部，以75%功率将该箱放入700瓦的微波炉中，使用8分钟。在治疗结束时，打开盖子，让溶液冷却到室温，然后用三个五分钟的新鲜 PBST 洗涤在科普林罐中洗涤幻灯片。要阻止任何非特异性绑定站点，在上次洗涤后，从每张幻灯片中摇动多余的 PBST，并覆盖幻灯片，并提供足够的卷适当的阻塞解决方案。在加湿室的室温下30分钟后，摇动每个幻灯片中的过量阻断溶液，并添加250微升的首批原发抗体。为了防止样品干燥，滑梯可能覆盖着一块石蜡薄膜。在加湿室中4摄氏度下过夜孵育后，用新鲜PBST清洗幻灯片三次，每次洗涤5分钟。甩掉多余的PBST，然后用250微升的合适二次抗体溶液快速覆盖每张幻灯片，在室温下在加湿室中孵育一小时。在孵化结束时，如所证明，在新鲜的PBST中清洗幻灯片三次，并在每张幻灯片中加入250微升新鲜准备好的石勒普他丁马萝卜过氧化物。在室温下 30 分钟后，用新鲜的 PBST 清洗幻灯片三次。然后摆脱多余的PBST，用250微升新鲜准备好的氟磷-酪酰胺溶液覆盖每张幻灯片。一分钟后，将幻灯片淹没在新鲜的 PBST 中，然后将三次两分钟的洗涤淹没在新鲜的 PBST 中。上次洗涤后，将几滴一对一甘油对PBS溶液涂抹在各部分，通过荧光显微镜检查荧光信号是否存在。要剥离第一个抗体复合物，请将抗体标记的幻灯片放入275毫升沸腾的柠檬酸钠溶液中至少8分钟，如所证明的。在治疗结束时，打开盖子，让溶液冷却至室温，然后用 PBST 洗三次幻灯片，每次洗涤五分钟。为了染色的第二个目标蛋白感兴趣，在阻止非特异性结合后，标记每个样品与250微升的第二主要抗体感兴趣的在四摄氏度过夜。第二天早上，每次洗涤用新鲜 PBST 洗三次，洗涤幻灯片三次，每次洗涤时用 250 微升的合适二次抗体溶液覆盖每个幻灯片，在室温下孵育一小时。在孵化结束时，如所证明，在新鲜的PBST中清洗幻灯片三次，并在每张幻灯片中加入250微升新鲜准备好的石勒普他丁马萝卜过氧化物。为了开发感兴趣的第二个目标蛋白的信号，经过三次PBST洗涤后，用荧光光谱中与氟磷结合的酪酰胺处理幻灯片。要停止泰酰胺信号的发展，请将幻灯片淹没到 PBST 中。然后用适当的核染料染色样品。对于荧光显微镜成像，在盖玻片中安装每个标有核染料的幻灯片，并放置一滴适当的安装介质。使用 4X 目标定位幻灯片上的组织。切换到 10 倍目标进行成像，将曝光时间调整为 100 至 400 毫秒，光源的强度在 40 到 80% 之间。然后获取每个通道中的图像，而无需移动舞台。每个通道的图像可以合并为多路复用荧光图像。在不剥离第一抗体染色和二次抗体染色之间，酪氨酸羟基酶的第二染色将从感兴趣的第一个目标蛋白中接收信号，从而产生假阳性酪氨酸羟基酶染色，例如在肾上腺皮质内的这个样本中。为了从第一次免疫染色中去除马萝卜过氧化物酶中的抗体交叉反应，可以进行1分钟和8分钟的微波中位剥离，从而产生干净的双重染色结果。在负对样本中未拾取任何显著信号。在沸腾的柠檬酸盐缓冲液中剥离8分钟也足以消除肾上腺中常用的许多抗体的抗体交叉反应。然而，在某些情况下，弱误报信号仍然可检测到，微波治疗增加20分钟仍可能无法完全消除抗体交叉反应。在荧光成像过程中，在切换通道时不要移动舞台非常重要。但是，可能需要重新调整焦点，以获得每个通道的清晰图像。对于每个通道，请务必调整曝光时间，以确保信号适当曝光，而图像中没有过度曝光的像素或区域。在染色过程中，保持滑轨从脱蜡中水分一部分非常重要，并且可以使用相同的剥离方法多次剥离幻灯片，以允许多路复用染色。

Research

JoVE Journal

Developmental Biology

Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals &#8722; Using Unconjugated Primary Antibodies from the Same Host Species

Production and Use of Lentivirus to Selectively Transduce Primary Oligodendrocyte Precursor Cells for In Vitro Myelination Assays

Production and Use of Lentivirus to Selectively Transduce Primary Oligodendrocyte Precursor Cells for In Vitro Myelination Assays

生产和使用慢病毒的有选择性地转导原少突胶质前体细胞<em&gt;在体外</em&gt;髓鞘测定

Myelination is a complex process that involves both neurons and the myelin forming glial cells, oligodendrocytes in the central nervous system (CNS) and ...

科研

发育生物学

Alginate Encapsulation of Pluripotent Stem Cells Using a Co-axial Nozzle

多能干细胞的海藻酸钠封装使用同轴喷嘴

Pluripotent stem cells (PS cells) are the focus of intense research due to their role in regenerative medicine and drug screening.  However, the ...

Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements

使用分离线粒体从小鼠骨骼肌进行高通量微孔板呼吸测量最小数量

Skeletal muscle mitochondria play a specific role in many disease pathologies. As such, the measurement of oxygen consumption as an indicator of ...

Generating Primary Fibroblast Cultures from Mouse Ear and Tail Tissues

从鼠耳和尾组织生成主成纤维细胞培养

Primary cells are derived directly from tissue and are thought to be more representative of the physiological state of cells in vivo than established cell ...

Induced Pluripotent Stem Cell Generation from Blood Cells Using Sendai Virus and Centrifugation

从血细胞使用仙台病毒和离心诱导多能干细胞代

The recent development of human induced pluripotent stem cells (hiPSCs) proved that mature somatic cells can return to an undifferentiated, pluripotent ...

Using Primary Neurosphere Cultures to Study Primary Cilia

使用主神经球文化研究纤毛

The primary cilium is fundamentally important for the proliferation of neural stem/progenitor cells and for neuronal differentiation during embryonic, ...

Whole-cell Patch-clamp Recordings of Isolated Primary Epithelial Cells from the Epididymis

分离的原发性上皮细胞从附睾的全细胞贴片钳记录

The epididymis is an essential organ for sperm maturation and reproductive health. The epididymal epithelium consists of intricately connected cell types ...

Genetic Engineering of Primary Mouse Intestinal Organoids Using Magnetic Nanoparticle Transduction Viral Vectors for Frozen Sectioning

使用磁纳米粒子转导病毒载体进行冷冻切片的原小鼠肠道器官的遗传工程

Intestinal organoid cultures provide a unique opportunity to investigate intestinal stem cell and crypt biology in vitro, although efficient approaches to ...

Induction and Validation of Cellular Senescence in Primary Human Cells

原发性细胞衰老的诱导与鉴定

Cellular senescence is a state of permanent cell cycle arrest activated in response to different damaging stimuli. Activation of cellular senescence is a ...

Tissue Preparation and Immunostaining of Mouse Craniofacial Tissues and Undecalcified Bone

小鼠颅面组织和未切除骨骼的组织制备及免疫染色

Tissue immunostaining provides highly specific and reliable detection of proteins of interest within a given tissue. Here we describe a complete and ...