Name: live cell imaging to assess the dynamics of metaphase timing and cell fate following mitotic spindle perturbations
Uploaded: 2019-09-20T15:00:00
Description: Watch this Scientific Journal Video about Live Cell Imaging to Assess the Dynamics of Metaphase Timing and Cell Fate Following Mitotic Spindle Perturbations at JoVE.com

DLM 由 NSF GRFP 支持。ALM 由史密斯家族生物医学研究卓越奖资助。

1. 生成 hTERT-RPE-1 细胞，可稳稳地表达 RFP-组蛋白 2B （RFP-H2B） 和 β-图布林-EGFP （tub-EGFP）
注：所有步骤均采用无菌技术，在生物安全 II+ （BSL2+） 安全柜中执行。
<ol><li>根据制造商对脂质转染输送系统的说明，通过转染293T细胞，使用适当的慢病毒质粒，生成携带相关基因（β-图布林-EGFP和RFP-H2B）的逆转录病毒。<ol>
<li>第1天：使用一次性玻璃巴斯德移液器从一盘亚汇合293T细?...

<ol>
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D., Degrassi, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Merotelic+kinetochore+orientation+versus+chromosome+mono-orientation+in+the+origin+of+lagging+chromosomes+in+human+primary+cells.">Merotelic kinetochore orientation versus chromosome mono-orientation in the origin of lagging chromosomes in human primary cells.</a> Journal of Cell Science. 115, 507-515 (2002).</li><li>Kwon, 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=Mechanisms+to+suppress+multipolar+divisions+in+cancer+cells+with+extra+centrosomes.">Mechanisms to suppress multipolar divisions in cancer cells with extra centrosomes.</a> Genes &amp; Development. 22, 2189-2203 (2008).</li><li>Khodjakov, A., Rieder, C. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aurora+A+inhibition+limits+centrosome+clustering+and+promotes+mitotic+catastrophe+in+cells+with+supernumerary+centrosomes.">Aurora A inhibition limits centrosome clustering and promotes mitotic catastrophe in cells with supernumerary centrosomes.</a> Oncotarget. 10, 1649-1659 (2019).</li><li>Conte, N., 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=TACC1-chTOG-Aurora+A+protein+complex+in+breast+cancer.">TACC1-chTOG-Aurora A protein complex in breast cancer.</a> Oncogene. 22, 8102-8116 (2003).</li><li>Schumacher, J. M., Ashcroft, N., Donovan, P. J., Golden, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+highly+conserved+centrosomal+kinase,+AIR-1,+is+required+for+accurate+cell+cycle+progression+and+segregation+of+developmental+factors+in+Caenorhabditis+elegans+embryos.">A highly conserved centrosomal kinase, AIR-1, is required for accurate cell cycle progression and segregation of developmental factors in Caenorhabditis elegans embryos.</a> Development. 125, 4391-4402 (1998).</li><li>Asteriti, I. A., Giubettini, M., Lavia, P., Guarguaglini, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aurora-A+inactivation+causes+mitotic+spindle+pole+fragmentation+by+unbalancing+microtubule-generated+forces.">Aurora-A inactivation causes mitotic spindle pole fragmentation by unbalancing microtubule-generated forces.</a> Molecular Cancer. 10, 131 (2011).</li><li>Chan, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+clinical+overview+of+centrosome+amplification+in+human+cancers.">A clinical overview of centrosome amplification in human cancers.</a> International Journal of Biological Sciences. 7, 1122-1144 (2011).</li><li>Kramer, A., Maier, B., Bartek, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Centrosome+clustering+and+chromosomal+(in)stability:+a+matter+of+life+and+death.">Centrosome clustering and chromosomal (in)stability: a matter of life and death.</a> Molecular Oncology. 5, 324-335 (2011).</li><li>Ganem, N. J., Godinho, S. A., Pellman, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mechanism+linking+extra+centrosomes+to+chromosomal+instability.">A mechanism linking extra centrosomes to chromosomal instability.</a> Nature. 460, 278-282 (2009).</li><li>Silkworth, W. T., Nardi, I. K., Scholl, L. M., Cimini, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multipolar+spindle+pole+coalescence+is+a+major+source+of+kinetochore+mis-attachment+and+chromosome+mis-segregation+in+cancer+cells.">Multipolar spindle pole coalescence is a major source of kinetochore mis-attachment and chromosome mis-segregation in cancer cells.</a> PLoS One. 4, e6564 (2009).</li><li>Nigg, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Centrosome+aberrations:+cause+or+consequence+of+cancer+progression?.">Centrosome aberrations: cause or consequence of cancer progression?.</a> Nature reviews, Cancer. 2, 815-825 (2002).</li><li>Godinho, S. A., Kwon, M., Pellman, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Centrosomes+and+cancer:+how+cancer+cells+divide+with+too+many+centrosomes.">Centrosomes and cancer: how cancer cells divide with too many centrosomes.</a> Cancer Metastasis Reviews. 28, 85-98 (2009).</li><li>Quintyne, N. J., Reing, J. E., Hoffelder, D. R., Gollin, S. M., Saunders, W. 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延时成像提供的时间分辨率允许可视化和评估单个细胞内的顺序细胞事件。利用细胞同步，随后在连续时间点收集和固定细胞的方法是有限的，因为最终比较是在细胞群之间进行的。在细胞对扰动的反应可能不均匀，或者可视化过程是动态的环境下，活细胞延时成像更有能力跟踪和分析单个细胞的动态以及异质性在细胞群中。通过这种方式，延时成像在监测细胞通过线粒体分裂的动态阶段的进展?...

基于图像的固定细胞分析通常用于评估细胞总体水平的变化，以响应各种扰动。当与细胞同步结合，然后采集和成像串行时间点时，这些方法可用于建议细胞事件序列。然而，固定细胞成像是有限的，因为时间关系是隐含在一个群体，而不是在单个细胞的水平证明。这样，虽然固定细胞成像和分析足以观察强健的表型和稳定状态变化，但检测随时间变化和仅影响细胞亚群的变化的能力并不完美。相反，活细胞成像是一个雄辩的工具，可用于观察单个细胞或细胞群中的细胞和亚细胞过程，随着时间的推移，无需辅助同步方法，这些方法本身可能会影响细胞行为1,2,3,4,5,6.
双相线轴的形成对于细胞分裂期间适当的染色体分离至关重要，从而产生两个基因相同的子细胞。线粒主轴结构的缺陷会破坏线粒体进展，并损害染色体分离的保真度，从而导致灾难性细胞分裂和细胞生存能力下降。因此，改变主轴形成的线粒体毒素是限制癌细胞7、8、9快速增殖的有希望的治疗方法。然而，在添加线粒毒物后，主轴结构的固定细胞分析在评估主轴形成的动态过程的能力方面受到限制，并且可能不能指示观察到的主轴结构变化是永久性的还是相反是暂时的，可以克服，以允许成功的细胞分裂。
在此协议中，我们描述了一种通过活细胞成像评估主轴扰动后线粒体动力学的方法。使用 hTERT 不朽的 RPE-1 细胞系，用于表达 RFP 标记的 Histone 2B 来可视化染色质，以及 EGFP 标记的 α-tubulin 来可视化微管、元相染色体对齐的时间、抗癫痫的发病，并最终线粒细胞的命运是使用染色体运动，压实和核形态的视觉线索评估。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.05% Trypsin</td>
			<td>Gibo-Life sciences</td>
			<td>25-510</td>
			<td>A serine protease used to release adherent cells from culture dishes</td>
		</tr>
		<tr>
			<td>15ml centrifuge tubes</td>
			<td>Olympus Plastics</td>
			<td>28-101</td>
			<td></td>
		</tr>
		<tr>
			<td>20x CFI Plan Fluor objective&#160;</td>
			<td>Nikon</td>
			<td></td>
			<td>For use in Live-cell imaging to visualize both bright field and fluorescence</td>
		</tr>
		<tr>
			<td>293T Cells</td>
			<td>ATCC</td>
			<td>CRL-3216</td>
			<td>For use in retroviral transfection; used in step 1.1</td>
		</tr>
		<tr>
			<td>a-tubulin-EGFP&#160;</td>
			<td>Addgene</td>
			<td>various numbers</td>
			<td>Expression vector for alpha tubulin fused to a green fluorescent protein tag; for use in the visualization of tubulin in live-cell imaging: commercially available through addgene and other vendors</td>
		</tr>
		<tr>
			<td>Alisertib</td>
			<td>Selleckchem</td>
			<td>S1133</td>
			<td>Small molecule inhibitor of the mitotic kinase Aurora A. Stock concentration is prepared at 10mM in DMSO, and used at a final concentration of 100nM.</td>
		</tr>
		<tr>
			<td>Blasticidin</td>
			<td>Invitrogen</td>
			<td>A11139-03</td>
			<td>Antibiotic selection agent; used to select for a-tub-EGFP expressing cells</td>
		</tr>
		<tr>
			<td>C02</td>
			<td>Airgas</td>
			<td></td>
			<td>For use in cell culture and live cell imaging</td>
		</tr>
		<tr>
			<td>Chroma ET-DS Red (TRITC/Cy3)</td>
			<td>Chroma</td>
			<td>49005</td>
			<td>Single band filter set; excitation wavelength 545nm with 25nm bandwidth and emission at 605nm wavelength with 70nm bandwidth; for visualization of H2B-RFP</td>
		</tr>
		<tr>
			<td>Chroma ET-EGFP (FITC/Cy2)</td>
			<td>Chroma</td>
			<td>49002</td>
			<td>Single band filter set; excitation wavelength 470nm with 40nm bandwidth and emission at 525nm wavelength with 50nm bandwidth; for visualization of GFP-tubulin</td>
		</tr>
		<tr>
			<td>disposable glass Pastuer pipets, sterilized&#160;</td>
			<td>Fisher Scientific</td>
			<td>13-678-6A</td>
			<td>For use in aspirating cells&#160;</td>
		</tr>
		<tr>
			<td>Dulbecco&#8217;s Modified Eagle Medium (DMEM)</td>
			<td>Gibo-Life sciences</td>
			<td>11965-084</td>
			<td>Cell culture medium for growth of RPE-1 and 293T cells</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum (FBS)</td>
			<td>Gibo-Life sciences</td>
			<td>10438-026</td>
			<td>Cell culture medium supplement</td>
		</tr>
		<tr>
			<td>Lipofectamine 3000 and p3000</td>
			<td>Invitrogen</td>
			<td>L3000-015</td>
			<td>Lipid based transfection reagent for transfection of plasmids; used in 1.1.4</td>
		</tr>
		<tr>
			<td>Multi well Tissue Culture dishes</td>
			<td>Corning</td>
			<td>various</td>
			<td>for use in cell culture, transfection/infection, and live cell imaging</td>
		</tr>
		<tr>
			<td>Nikon Ti-E microscope</td>
			<td>Nikon</td>
			<td></td>
			<td>Inverted epifluorescence microscope for use in live-cell imaging</td>
		</tr>
		<tr>
			<td>NIS Elements HC&#160;</td>
			<td>Nikon</td>
			<td>Version 4.51</td>
			<td>Image acquisition and analysis software; used in sections 3 &#38; 4</td>
		</tr>
		<tr>
			<td>OPTI-MEM</td>
			<td>Gibo-Life sciences</td>
			<td>31985-070</td>
			<td>Reduced serum medium for cell transfection; used in step 1.1.3</td>
		</tr>
		<tr>
			<td>Penicillin/Streptomycin&#160;</td>
			<td>Gibo-Life sciences</td>
			<td>15140-122</td>
			<td>antibiotic used in cell culture medium</td>
		</tr>
		<tr>
			<td>phosphate bufferred saline (PBS)</td>
			<td>Caisson labs</td>
			<td>PBP06-10X1LT</td>
			<td>sterile saline solution for use with cell culture</td>
		</tr>
		<tr>
			<td>pMD2.G</td>
			<td>Addgene</td>
			<td>12259</td>
			<td>Lentiviral VSV-G envelope expression construct; used in step 1.1.4</td>
		</tr>
		<tr>
			<td>Polybrene</td>
			<td>Sigma-Aldrich</td>
			<td>H9268</td>
			<td>Cationic polymer used to enhane viral infection efficiency; used in step 1.1.10</td>
		</tr>
		<tr>
			<td>psPAX</td>
			<td>Addgene</td>
			<td>12260</td>
			<td>2nd generation lentiviral packaging plasmid; used in step 1.1.4</td>
		</tr>
		<tr>
			<td>Puromycin</td>
			<td>Invitrogen</td>
			<td>ant-pr-1</td>
			<td>Antibiotic selection agent; used to select for RFP-H2B expressing cells</td>
		</tr>
		<tr>
			<td>RFP- Histone 2B (H2B)</td>
			<td>Addgene</td>
			<td>various numbers</td>
			<td>Expression vector for red fluorescent protein-tagged histone 2B; for use in the visualization of chromatin in live-cell imaging: commercially available through Addgene and other vendors</td>
		</tr>
		<tr>
			<td>RNAi Max</td>
			<td>Invitrogen</td>
			<td>13778-150</td>
			<td>Lipid based transfection reagent for transfection of siRNA constructs</td>
		</tr>
		<tr>
			<td>RPE-1 cells</td>
			<td>ATCC</td>
			<td>CRL-4000</td>
			<td>Human retinal pigment epithelial cell line</td>
		</tr>
		<tr>
			<td>Tissue culture dish 100x20mm</td>
			<td>Corning&#160;</td>
			<td>353003</td>
			<td>for use in culturing adherent cells</td>
		</tr>
		<tr>
			<td>Zyla sCMOS camera&#160;</td>
			<td>Nikon</td>
			<td></td>
			<td>Camera attached to the micrscope, used for capturing images of cells</td>
		</tr>
	</tbody>
</table>

live cell imaging to assess the dynamics of metaphase timing and cell fate following mitotic spindle perturbations

活细胞延时成像是细胞生物学中的重要工具，它提供了对细胞过程的洞察，否则这些细胞过程可能被固定细胞分析忽略、误解或误解。虽然固定细胞成像和分析是健壮的，足以观察细胞稳定状态，但它在定义细胞级事件的时间顺序时可能受到限制，并且无法评估动态过程的瞬态性质，包括线粒体进展。相反，活细胞成像是一种雄辩的工具，可用于观察细胞过程在单细胞水平随着时间的推移，并有能力捕捉过程的动态，否则在固定细胞成像中表现不佳。在这里，我们描述了一种生成带有荧光标记染色质和微管标记的细胞的方法，以及它们在活细胞成像方法中的使用，以监测元相染色体对齐和线粒出口。我们描述了基于成像的技术，以评估主轴形成和线粒进展的动态，包括识别线粒体不同阶段的细胞，识别和跟踪线粒缺陷，以及分析主轴动力学和使用线粒抑制剂治疗后的线粒细胞命运。

在主轴扰动存在时线粒体进展的评估 主轴聚焦的调节是正确双极主轴形成的关键步骤。通过蛋白质耗尽、药物抑制或中心体数改变破坏这一过程，破坏主轴结构，延迟或停止线粒体进展10、11、12、13。然而，一些扰动只是暂时延迟主轴的形成与细胞最终通过线?...

Watch this Scientific Journal Video about Live Cell Imaging to Assess the Dynamics of Metaphase Timing and Cell Fate Following Mitotic Spindle Perturbations at JoVE.com

Live Cell Imaging to Assess the Dynamics of Metaphase Timing and Cell Fate Following Mitotic Spindle Perturbations

在这里，我们提出一个协议，以评估主轴形成和线粒体进展的动态。我们的延时成像应用使用户能够识别线粒体不同阶段的细胞，跟踪和识别线粒体缺陷，并在接触抗线粒药物时分析主轴动力学和线粒细胞命运。

活细胞成像，以评估在斜接主轴扰动后元相定时和细胞命运的动态

Live cell time-lapse imaging is an important tool in cell biology that provides insight into cellular processes that might otherwise be overlooked, ...

活细胞成像可在单个细胞内观察动态细胞过程。这些方法对于评估影响细胞分裂动态过程的条件以及揭示分粒体缺陷对子细胞增殖和生存能力的影响是强大的。成像过程中光暴露导致的光毒性是与此协议相关的重大挑战。因此，应优化曝光时间、成像间隔和持续时间。使用无菌技术和生物安全二级安全柜，使用无菌一次性玻璃巴斯德移液器从含有含有具有兴趣表达构造的细胞系的培养板中吸出介质。用10毫升无菌PBS用旋转来短暂清洗细胞，用2毫升0.05%的三辛辛对细胞进行治疗。在37摄氏度下2至5分钟后，在板中加入8毫升新鲜介质，停止反应，用温和的移液重新暂停分离的细胞。将细胞溶液转移到无菌的15毫升管中，通过离心收集细胞。将颗粒重新在 10 毫升 PBS 中重新浓缩，然后再次使细胞离心。将颗粒重新在10毫升的培养基中进行计数，将细胞稀释至每毫升细胞培养基浓度的1至2倍至第5细胞。然后将500微升细胞播种到无菌12井成像底板的每个井中，将该细胞放入细胞培养箱，使细胞粘附在细胞表面。成像前不超过30分钟，在一口或多口井中加入相关浓度的线粒体药物，并将抑制剂稀释剂的等量添加到与对口的相同数量的井中。为准备成像显微镜，请将板放在装有高分辨率摄像机的倒置荧光显微镜的舞台上，将环境室预热至 37 摄氏度，并安装加湿 5%二氧化碳的输送系统。选择 20 倍空气目标，数值孔径为 0.5，并配备用于高对比度荧光和相位对比度，或明亮的场成像和查看细胞以调整过程并找到焦点以聚焦细胞。要设置亮场、GFP 和 RFP 图像采集的最佳曝光时间，请选择相应的滤波块，并为每个将成像的荧光团选择相应的激发和发射，然后单击"播放"。如果信号不够强烈，请调整自动曝光，并在每个荧光通道中从"格式"下拉菜单中选择适当的装箱。根据制造商的说明选择显微镜阶段并校准到多孔盘，并使用图像采集软件突出显示或以其他方式选择要成像的孔。打开生成的点控制面板，在"工作区"下选择"受限"和"边界"以限制坐标选择区域以排除井的边界。在"区域限制"下拉菜单中，选择"整个区域"并选择随机点放置。将计数设置为 6，然后单击"随机化"，分别选择每个井捕获的点的数量和分布。然后单击并在时间序列控制面板中输入值，以选择并输入收集图像的时间间隔和持续时间。要可视化标有 RFP-H2B 的染色质，请选择用 RFP 滤波块拍摄的图像，并识别进入线粒体（如初始染色质压实和核包络分解所示）的细胞，以确定单个细胞中元相对齐和肛度发作的线粒体时间。通过采集的影片中的连续时间点跟踪感兴趣的细胞，以确定从线粒体进入到 RFP-H2B 标记的染色质在元相期间完成细胞赤道对齐的时间点数或分钟数。为了监测线粒体定时、线粒体保真度和细胞命运，请继续通过连续的时间点跟踪细胞，以确定相位染色体分离明显和/或染色质去编译和核包络改造发生的时间坐标。然后可视化 RFP-H2B，以识别每个群体中表现出线粒体缺陷的细胞，包括在相位染色体分离期间滞后的染色体和染色质桥。通过可视化α浴缸-EGFP，可以观察到，当主轴拉对焦实现并形成双极性线粒体主轴以准备细胞分裂时，经历主轴中断的细胞会发生动态变化。与主轴组装同时，染色体运动可以通过RFP-H2B进行可视化，以评估染色体对齐和分离保真度。利用活细胞成像方法，这些具有代表性的结果表明，具有正常中心体含量的细胞能够从核包络分解到元相对齐和离相发作，在30分钟内实现双极分裂。在额外的中体存在的情况下，近50%的细胞能够克服瞬态多极线线粒体主轴，并在完整的细胞分裂中形成双极主轴。剩余的细胞无法实现双极主轴，因此，通过多极分裂退出线粒体。无论是否实现主轴双极性，与具有两个中体细胞的细胞相比，具有额外中体细胞的分粒期显著增加。指示线粒体进展的动态可能会改变，即使线粒体结果的变化不明显。在定义荧光通道的曝光时间时，优化曝光时间以限制光毒性。此外，还可以选择摄像机像素装箱，以便使用更低的曝光时间。此过程适用于药理筛选、入侵和迁移分析。这些方法也可以用于检查药物治疗的疗效或细胞活动动力学的动态。

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Education

JoVE Journal

JoVE Core

Molecular Biology

Biology

Unit 2

Cell Proliferation

SCIENTISTS IN ACTION

Live Cell Imaging of F-actin Dynamics via Fluorescent Speckle Microscopy (FSM)

Live Cell Imaging of F-actin Dynamics via Fluorescent Speckle Microscopy FSM

F - actin的动力学通过荧光活细胞成像斑点显微镜（密克罗尼西亚）

In this protocol we describe the use of Fluorescent Speckle Microscopy (FSM) to capture high-resolution images of actin dynamics in PtK1 cells. A unique ...

科研

生物学

Assembly, Loading, and Alignment of an Analytical Ultracentrifuge Sample Cell

大会，加载和分析超速离心机样品池对齐

The analytical ultracentrifuge (AUC) is a powerful biophysical tool that allows us to record macromolecular sedimentation profiles during high speed ...

Single Cell Fate Mapping in Zebrafish

单细胞在斑马鱼的命运映射

The ability to differentially label single cells has important implications in developmental biology. For instance, determining how hematopoietic, ...

Blastomere Explants to Test for Cell Fate Commitment During Embryonic Development

卵裂球外植体测试胚胎发育过程中细胞命运的承诺

Fate maps, constructed from lineage tracing all of the cells of an embryo, reveal  which tissues descend from each cell of the embryo.  Although fate maps ...

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis

凋亡细胞清除过程中实时成像<em&gt;果蝇</em&gt;胚胎发育

The  proper elimination of unwanted or aberrant cells through apoptosis and  subsequent phagocytosis (apoptotic cell clearance) is crucial for normal  ...

Live Cell Imaging of Early Autophagy Events: Omegasomes and Beyond

活细胞成像早期自噬活动：Omegasomes与超越

Autophagy is a  cellular response triggered by the lack of nutrients, especially the absence of  amino acids. Autophagy is defined by the formation of ...

Live Cell Imaging of Primary Rat Neonatal Cardiomyocytes Following Adenoviral and Lentiviral Transduction Using Confocal Spinning Disk Microscopy

原代大鼠乳鼠心肌细胞的活细胞成像继腺病毒和慢病毒转导利用共聚焦旋转盘显微镜

Primary rat neonatal cardiomyocytes are useful in basic in vitro cardiovascular research because they can be easily isolated in large numbers in a single ...

Studying the Protein Quality Control System of D. discoideum Using Temperature-controlled Live Cell Imaging

Studying the Protein Quality Control System of D. discoideum Using Temperature-controlled Live Cell Imaging

研究蛋白质的质量控制体系<em&gt;Ð。菌</em&gt;使用温控活细胞成像

The complex lifestyle of the social amoebae Dictyostelium discoideum makes it a valuable model for the study of various biological processes. Recently, we ...

Examination of Mitotic and Meiotic Fission Yeast Nuclear Dynamics by Fluorescence Live-cell Microscopy

荧光活细胞显微镜对微孔裂变酵母核动力学的检测

Live-cell imaging is a microscopy technique used to examine cell and protein dynamics in living cells. This imaging method is not toxic, generally does ...

Super-Resolution Live Cell Imaging of Subcellular Structures

亚细胞结构的超分辨率活细胞成像

There has long been a crucial tradeoff between spatial and temporal resolution in imaging. Imaging beyond the diffraction limit of light has traditionally ...