Name: measurement of mitochondrial mass and membrane potential in hematopoietic stem cells and t-cells by flow cytometry
Uploaded: 2019-12-26T14:00:00
Description: Watch this Scientific Journal Video about Measurement of Mitochondrial Mass and Membrane Potential in Hematopoietic Stem Cells and T-cells by Flow Cytometry at JoVE.com

我们感谢联伊会流动细胞学核心设施的支持，特别是罗曼·贝德尔博士的支持。这项工作得到了克里斯蒂安·格哈德·杰布森基金会对N.V.和O.N.基金会的资助。

手稿中描述的所有实验均遵循我们机构的指导方针，并按照瑞士动物实验法（授权：VD3194）和涉及人类样本的研究（协议：235/14;CER-VD 2017-00490）
1. 造血干细胞提取
<ol><li>购买野生型C57BL6/J小鼠，并将其保存在动物之家至少一周，以减少运输相关的压力。</li>
	<li>在实验当天，使用CO2窒息对小鼠实施安乐死。</li>
	<li>用70%乙醇喷洒小鼠，对毛皮进行消毒，并使用标准手?...

<ol>
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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=Durable+complete+responses+in+heavily+pretreated+patients+with+metastatic+melanoma+using+T+cell+transfer+immunotherapy.">Durable complete responses in heavily pretreated patients with metastatic melanoma using T cell transfer immunotherapy.</a> Clinical Cancer Research. 17 (13), 4550-4557 (2011).</li><li>Aranda, F., 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=Trial+Watch:+Adoptive+cell+transfer+for+anticancer+immunotherapy.">Trial Watch: Adoptive cell transfer for anticancer immunotherapy.</a> Oncoimmunology. 3, 28344 (2014).</li><li>Wherry, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=T+cell+exhaustion.">T cell exhaustion.</a> Nature Immunology. 12 (6), 492-499 (2011).</li><li>Ho, P. C., 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=Phosphoenolpyruvate+Is+a+Metabolic+Checkpoint+of+Anti-tumor+T+Cell+Responses.">Phosphoenolpyruvate Is a Metabolic Checkpoint of Anti-tumor T Cell Responses.</a> Cell. 162 (6), 1217-1228 (2015).</li><li>Chang, C. H., 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=Metabolic+Competition+in+the+Tumor+Microenvironment+Is+a+Driver+of+Cancer+Progression.">Metabolic Competition in the Tumor Microenvironment Is a Driver of Cancer Progression.</a> Cell. 162 (6), 1229-1241 (2015).</li><li>van der Waart, A. 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=Inhibition+of+Akt+signaling+promotes+the+generation+of+superior+tumor-reactive+T+cells+for+adoptive+immunotherapy.">Inhibition of Akt signaling promotes the generation of superior tumor-reactive T cells for adoptive immunotherapy.</a> Blood. 124 (23), 3490-3500 (2014).</li><li>Crompton, J. 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=Akt+inhibition+enhances+expansion+of+potent+tumor-specific+lymphocytes+with+memory+cell+characteristics.">Akt inhibition enhances expansion of potent tumor-specific lymphocytes with memory cell characteristics.</a> Cancer Research. 75 (2), 296-305 (2015).</li><li>de Almeida, M. J., Luchsinger, L. L., Corrigan, D. J., Williams, L. J., Snoeck, H. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dye-Independent+Methods+Reveal+Elevated+Mitochondrial+Mass+in+Hematopoietic+Stem+Cells.">Dye-Independent Methods Reveal Elevated Mitochondrial Mass in Hematopoietic Stem Cells.</a> Cell Stem Cell. 21 (6), 725-729 (2017).</li><li>Abe, T., 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=Establishment+of+Conditional+Reporter+Mouse+Lines+at+ROSA26+Locus+For+Live+Cell+Imaging.">Establishment of Conditional Reporter Mouse Lines at ROSA26 Locus For Live Cell Imaging.</a> Genesis. 49 (7), 579-590 (2011).</li><li>Sukumar, 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=Mitochondrial+Membrane+Potential+Identifies+Cells+with+Enhanced+Stemness+for+Cellular+Therapy.">Mitochondrial Membrane Potential Identifies Cells with Enhanced Stemness for Cellular Therapy.</a> Cell Metabolism. 23 (1), 63-76 (2016).</li><li>Sukumar, 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=Inhibiting+glycolytic+metabolism+enhances+CD8++T+cell+memory+and+antitumor+function.">Inhibiting glycolytic metabolism enhances CD8+ T cell memory and antitumor function.</a> Journal of Clinical Investigation. 123 (10), 4479-4488 (2013).</li></ol>

严格调节HSC功能对于维持生物体寿命期间稳定的造体非常重要。与体内其他各种细胞类型一样，促进HSC功能调节的一个关键成分是细胞代谢。我们实验室9和其他2，3的研究已经暗示了线粒体在HSCs中维持一种独特的代谢状态的重要性。 由于从鼠骨髓中分离出的HSCs数量极少，很难通过标准代谢检测（例如，用SeaHorse消耗氧气）来分析它们...

造血干细胞 （HSCs） 是存在于骨髓中的少量细胞，可确保在生物体的整个生命周期内产生血液和平衡。HSC通过产生后代来调解这个过程，这些后代又通过几轮细胞分裂和精心编排的分化步骤1产生最终分化的成熟血细胞谱系。重要的是，HSC通过厌氧糖解产生能量。相反，更坚定和活跃的造血原体将新陈代谢转向线粒体代谢2，3，4。这种独特的代谢状态被认为可以保护HSC免受活性线粒体产生的活性氧物种（ROS）造成的细胞损伤，从而保持其长期体内功能5、6、7、8。直接测量 HSC 代谢状态具有挑战性，并且由于数量有限，通常吞吐量较低。在这里，我们描述了一种基于流式细胞学的测定，用于使用四甲基二甲胺甲基酯（TMRM）荧光和线粒体质量在HSC中使用绿色荧光线粒体染色（Mitotracker Green）对线粒体膜电位（μm）进行可靠测量。我们之前已经证明，低μm是高度纯化的HSCs9和代谢调制器的一个真正的功能标记，能够降低μm增强HSCs功能9，10。在这里，我们建议使用我们的方法在HSC线粒体分析作为策略，以确定能够提高HSC的长期血液重组潜力的新分子。
例如，我们证明，这种测定在接触维生素B3模拟烟酰胺（NR）时，能可靠地测量HSCμm的降低。因此，在我们最近发表的研究中，我们证明NR通过直接改善造血干细胞和祖细胞功能10，大力改善小鼠和人化小鼠系统的血液恢复后。考虑到25%的死亡率与移植后患者血液延迟和免疫恢复有关，这种代谢调节器的能力具有很大的临床价值。
此外，我们提供了证据，这种方法可用于表征人类T细胞的代谢特征和功能。近年来，采用自体肿瘤渗透淋巴细胞（TILs）的采用细胞疗法（ACT）的发展已成为某些类型的晚期癌症最有效的方法，其预后极不利（例如，转移性黑色素瘤，其中>50%的患者对治疗有反应，高达24%的患者有完全的回归）13。然而，具有足够抗肿瘤活性的TILs很难产生14。TILs在体外扩张期间经历的广泛增殖和刺激导致T细胞耗尽和衰老，从而显著削弱T细胞抗肿瘤反应15。重要的是，TL的抗肿瘤能力与其代谢16、17和旨在通过抑制PI3K/Akt途径调节新陈代谢的方法紧密相连，产生了令人鼓舞的结果18、19。因此，我们比较了从外周血单核细胞（PBMC）和患者衍生的TLS中提取的T细胞的μm，并表明与最终分化的TL相比，分化较少的PBMC衍生T细胞具有较低的μμm和线粒体质量。
我们设想，这种测定可用于识别新的代谢调节剂，通过βm的调制改善HSC和T细胞功能。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>5 mL FACS tubes</td>
			<td>Falcon</td>
			<td>352235</td>
			<td>Sample preparation</td>
		</tr>
		<tr>
			<td>96-U bottom plate</td>
			<td>Corning</td>
			<td>3799</td>
			<td>Cell culture</td>
		</tr>
		<tr>
			<td>AutoMACS pro separator</td>
			<td>Miltenyi Biotec</td>
			<td></td>
			<td>Automatic Cell separation</td>
		</tr>
		<tr>
			<td>BD FACS AriaIII</td>
			<td>Becton and Dickinson</td>
			<td></td>
			<td>Cell sorting</td>
		</tr>
		<tr>
			<td>BD IMag mouse hematopoietic progenitor cell enrichment kit</td>
			<td>BD</td>
			<td>558451</td>
			<td>Lineage depletion</td>
		</tr>
		<tr>
			<td>BD LSRII</td>
			<td>Becton and Dickinson</td>
			<td></td>
			<td>FACS acquisition machine</td>
		</tr>
		<tr>
			<td>BD-DIVA</td>
			<td>Becton and Dickinson</td>
			<td></td>
			<td>Acquisition software</td>
		</tr>
		<tr>
			<td>CD150 PE</td>
			<td>Biolegend</td>
			<td>115904</td>
			<td>Antibody staining mix</td>
		</tr>
		<tr>
			<td>CD150 PE-Cy5</td>
			<td>Biolegend</td>
			<td>115912</td>
			<td>Antibody staining mix</td>
		</tr>
		<tr>
			<td>CD48 PB</td>
			<td>Biolegend</td>
			<td>103418</td>
			<td>Antibody staining mix</td>
		</tr>
		<tr>
			<td>Centrifuge- 5810R</td>
			<td>Eppendorf</td>
			<td></td>
			<td>Centrifugation</td>
		</tr>
		<tr>
			<td>Ckit PeCy7</td>
			<td>Biolegend</td>
			<td>105814</td>
			<td>Antibody staining mix</td>
		</tr>
		<tr>
			<td>Flow jo</td>
			<td>FlowJo LLC</td>
			<td></td>
			<td>FACS Analysis software</td>
		</tr>
		<tr>
			<td>GraphPad-Prism</td>
			<td>GraphPad</td>
			<td></td>
			<td>Plotting data into graphs</td>
		</tr>
		<tr>
			<td>Mitotracker Green</td>
			<td>Invitrogen</td>
			<td>M7514</td>
			<td>Green-fluorescent mitocondrial stain to measure mitochondrial mass; working concentration = 100 nM; stock concentration = 1 mM</td>
		</tr>
		<tr>
			<td>Nicotinamide Riboside (NR)</td>
			<td>Custom synthesized in house</td>
			<td></td>
			<td>Metabolic modulator; working concentration = 500 &#181;M; stock concentration = 50 mM</td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>CHUV</td>
			<td>1000324</td>
			<td>Buffer preparation; working concentration = 1x; stock concentration = 1x</td>
		</tr>
		<tr>
			<td>Pen-Strep (P/S)</td>
			<td>Life technologies</td>
			<td>15140122</td>
			<td>Ex vivo culture; working concentration = 1x; stock concentration = 1x</td>
		</tr>
		<tr>
			<td>RBC Lysis buffer</td>
			<td>Biolegend</td>
			<td>420301</td>
			<td>Lysing Red blood cells; working concentration = 1x; stock concentration = 10x</td>
		</tr>
		<tr>
			<td>Recombinant Mouse Flt-3 Ligand (FLT3)</td>
			<td>RnD</td>
			<td>427-FL-005/CF</td>
			<td>Ex vivo culture; working concentration = 2 ng/mL; stock concentration = 10 &#181;g/mL</td>
		</tr>
		<tr>
			<td>Recombinant mouse stem cell factor (SCF)</td>
			<td>RnD</td>
			<td>455-MC-010/CF</td>
			<td>Ex vivo culture; working concentration = 100 ng/mL; stock concentration = 50 &#181;g/mL</td>
		</tr>
		<tr>
			<td>Sca1 APC</td>
			<td>Thermo Fisher Scientific</td>
			<td>17-5981-82</td>
			<td>Antibody staining mix</td>
		</tr>
		<tr>
			<td>StemlineII Hematopoietic Stem Cell Expansion Medium</td>
			<td>SIGMA</td>
			<td>S0192</td>
			<td>Ex vivo culture</td>
		</tr>
		<tr>
			<td>Streptavidin Pac orange</td>
			<td>Life Technologies</td>
			<td>S32365</td>
			<td>Antibody staining mix</td>
		</tr>
		<tr>
			<td>Streptavidin Tx red</td>
			<td>Life Technologies</td>
			<td>S872</td>
			<td>Antibody staining mix</td>
		</tr>
		<tr>
			<td>TMRM</td>
			<td>Invitrogen</td>
			<td>T668</td>
			<td>Staining mitochondrial membrane potential; working concentration = 200 nM; stock concentration = 10 mM</td>
		</tr>
		<tr>
			<td>Ultra pure EDTA</td>
			<td>Invitrogen</td>
			<td>15575-038</td>
			<td>Buffer preparation; working concentration = 0.5 M; stock concentration = 1 mM</td>
		</tr>
	</tbody>
</table>

measurement of mitochondrial mass and membrane potential in hematopoietic stem cells and t-cells by flow cytometry

保持静止、自我更新和分化的精细平衡是保存造血干细胞（HSC）池和维持所有成熟血细胞的终身生产的关键。近年来，细胞代谢已成为HSC功能和命运的重要调节器。我们之前已经证明，线粒体代谢的调制会影响HSC的命运。具体来说，通过化学分离电子传输链，我们能够在通常诱导快速分化的培养条件下保持HSC功能。然而，限制HSC数通常阻止使用标准测定来测量HSC代谢，从而预测其功能。在这里，我们报告一个简单的流式细胞测定，允许可靠地测量在稀缺细胞（如HSC）中的线粒体膜电位和线粒体质量。我们讨论了从小鼠骨髓中分离HSC和线粒体质量的测量和膜电位在体外培养。例如，我们通过代谢调制器治疗，在HSC中显示了这些参数的调制。此外，我们扩展了这种方法在人类外周血源T细胞和人类肿瘤渗透淋巴细胞（TLL）中的应用，显示其线粒体特征的显著差异，可能反映不同的T细胞功能。我们相信，这种测定可用于筛选，以识别不同环境中不同细胞类型的线粒体代谢的调节剂。

在图1中，我们显示了从小鼠骨髓中分离造血干细胞的浇注策略及其体外培养板的布局。图 1A显示了 SSC-A/FSC-A 图中淋巴细胞分数的标识。在单门中取出双子细胞，然后由于没有DAPI信号，识别了活细胞。LKS种群，由血统-Sca1+cKit+定义，被识别。已知该种群含有干细胞和祖细胞。HSCs在LKS种群中形成大约5-10%的细胞，并通过CD150+ C...

Watch this Scientific Journal Video about Measurement of Mitochondrial Mass and Membrane Potential in Hematopoietic Stem Cells and T-cells by Flow Cytometry at JoVE.com

Measurement of Mitochondrial Mass and Membrane Potential in Hematopoietic Stem Cells and T-cells by Flow Cytometry

在这里，我们描述了一种可靠的方法来测量线粒体质量和膜电位在体外培养造血干细胞和T细胞。

流细胞测定方法测量造血干细胞和T细胞的线粒体质量及膜电位

A fine balance of quiescence, self-renewal, and differentiation is key to preserve the hematopoietic stem cell (HSC) pool and maintain lifelong production ...

该协议是一个简单的流细胞学为基础的测定，允许测量线粒体膜电位和线粒体质量在活细胞。标准代谢测定失败时，与罕见的人群，如造血干细胞。此方法使用户能够在使用低数量的细胞时进行线粒体膜分析，并确定新的细胞代谢调节器，如HSC。此外，您可以将它与下游功能分析（如骨髓移植或菌落形成分析）相结合，以确定培养后细胞的功能。我们的技术可用于识别在骨髓移植和造血疗法后造血免疫恢复的背景下能够改善HSC功能的新分子。如我们的协议所述，我们的方法可用于评估免疫T细胞的代谢健康。染色后，细胞对光线的暴露最小，这一点非常重要。此外，为了评估泵的回流的贡献，必须包括泵抑制剂维拉帕米尔的使用。然而，用户必须意识到，维拉帕米尔严重扰动离子平衡，不能用于评估代谢调节剂对线粒体膜电位的影响。此方法的可视化演示非常重要，因为在使用数量很少的单元格时，需要记住一些关键步骤，并且用户在 PubMed 上阅读的常规文件中经常缺少这些步骤。对于 FAC 排序，识别由血统负、Sca1 阳性、cKit 阳性定义的 LKS 总体。在LKS群体中，大约5至10%的细胞的造血干细胞通过CD150阳性、CD48阴性组分的浇注来识别，这些细胞被贴上HSC的标签。造血干细胞分选后，在摄氏4度下以300倍g离心管，5分钟。在不去除颗粒的情况下轻轻去除大部分上一提液，并在细胞颗粒上留下 50 到 80 微升。将干细胞扩张介质中的细胞颗粒重新用于最终体积为200微升。准备一个无菌组织培养处理96 U底井板，并确定将培养细胞的井。转移以前在这些油井中准备的 100 微升 2 倍基底介质。在 NR 标记良好，添加两微升的 100 倍烟酰胺核糖二聚二聚剂溶液。将 100 微升 2x 基底介质转移到孔中，用于染色控制。含有2倍基底介质的井顶部的已准备造血干细胞的种子100微升。种子100微升整个骨髓细胞在井标记为染色控制。在周围所有油井中加入200微升的自 <3> <3>化水，以减少含有细胞的井中的蒸发。将板在37摄氏度的培养箱中不受干扰地放置，在5%的二氧化碳中放置72小时。每24小时拿出一块盘子补充一次烟酰胺核糖酰胺，然后放回孵化器。在培养期结束时，在每个测试井中添加两个 100x TMRM 溶液的微升和两个 100 倍绿色荧光染色溶液的微升。在TMTM染色控制井中加入两个100倍TMTM微升。在 MitoTracker 染色控制井中加入两个 100 倍绿色荧光染色微升。用铝箔盖住板的顶部。将盘子放回孵化器中，在37摄氏度的20%二氧化碳中放置45分钟。将板从培养箱中取出，以 300 倍 g 离心 5 分钟。反转板以去除上流水，并添加 200 微升的标准 FACS 缓冲液重新暂停。用铝箔盖住盘子。以 300 次 g 离心板五分钟。重复此洗涤步骤三次。将细胞重新填充在 200 微升的 FACS 缓冲液缓冲器中，并转移到 FACS 管中。要运行流式圆环仪上的样本，请先将单个颜色控件加载到机器中，并逐个记录每个事件中的至少 5，000 个事件。在软件中，按"获取"，然后在仪表板上记录以获取和记录单个颜色控件。按"补偿设置"，然后单击"计算补偿"以计算补偿。一旦应用补偿，获取造血干细胞样本，并记录尽可能多的事件。从 Cytometer 导出 FACS 文件，并在分析软件上打开该文件。在 FlowJo 上，使用向前散射和侧散射门来标识细胞总体。识别下一个关卡中的单数，然后绘制表示活细胞的 DAPI 负分数。在活细胞门中，在TMM和绿色荧光染色通道中绘制一个轮廓图，分别测量线粒体膜电位和质量。在活细胞门中导出这两个通道的均荧光强度。然后，在所有样品中导出 TMRM 低门中的细胞比例。烟酰胺核糖酰胺治疗表明，TMRM低人口明显增加。它还显著增加了TMM低门中细胞的比例，并显示出TMTM荧光强度的显著降低。线粒体质量在烟酰胺核糖酰胺补充剂后没有改变。此外，干细胞标记染色与线粒体染色后培养相结合。通过识别造血干细胞的浇注策略，接触烟酰胺核糖酰胺表明TMM低人群显著增加，并显著降低了封闭HSC的TMM荧光强度。绿色荧光线粒体染色绿色信号在两个条件下保持不变。重要的是要用铝箔覆盖染色板，以尽量减少光线暴露在染色样品中。此方法可以与下游检测（如骨髓移植和菌落形成测定）相结合，以确定您的干细胞的功能。因此，该技术允许一个简单的筛选方法，用于识别HSC的新的代谢调节器。

Research

JoVE Journal

Biology

Isolation and Transplantation of Hematopoietic Stem Cells (HSCs)

Isolation and Transplantation of Hematopoietic Stem Cells HSCs

分离和造血干细胞移植（造血干细胞）

科研

生物学

Quantitative Measurement of GLUT4 Translocation to the Plasma Membrane by Flow Cytometry

流式细胞仪定量测定GLUT4转位到质膜

Glucose is the main source of energy for the body, requiring constant regulation of its blood concentration. Insulin release by the pancreas induces ...

Flow Cytometry Purification of Mouse Meiotic Cells

流动小鼠减数分裂细胞的流式细胞仪净化

The heterogeneous nature of cell types in the testis and the absence of meiotic cell culture models have been significant hurdles to the study of the ...

Phenotypic Analysis and Isolation of Murine Hematopoietic Stem Cells and Lineage-committed Progenitors

表型分析和分离小鼠造血干细胞和传承的承诺祖

The bone marrow is the principal site where HSCs and more mature blood cells lineage progenitors reside and differentiate in an adult organism. HSCs ...

Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes

优化细胞分裂监测细胞示踪染料的染色和扩散建模方法

Fluorescent cell tracking dyes, in combination with flow and image cytometry, are powerful tools with which to study the interactions and fates of ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

在造血干细胞和祖细胞的成年小鼠颅骨骨髓体内四维跟踪

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Cytosolic Calcium Measurements in Renal Epithelial Cells by Flow Cytometry

在肾小管上皮细胞胞浆钙的测量流式细胞仪

A variety of cellular processes, both physiological and pathophysiological, require or are governed by calcium, including exocytosis, mitochondrial ...

High Efficiency Differentiation of Human Pluripotent Stem Cells to Cardiomyocytes and Characterization by Flow Cytometry

人类多能干细胞的高效分化为心肌细胞，并鉴定流式细胞仪

There is an urgent need to develop approaches for repairing the damaged heart, discovering new therapeutic drugs that do not have toxic effects on the ...

Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy

线粒体钙和线粒体膜的同时测量通过荧光显微镜活细胞的潜在

Apart from their essential role in generating ATP, mitochondria also act as local calcium (Ca2+) buffers to tightly regulate intracellular Ca2+ ...

Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry

通过光谱流式细胞仪由固态组织中分离出悬浮细胞系的分析

Flow cytometry has been used for the past 40 years to define and analyze the phenotype of lymphoid and other hematopoietic cells. Initially restricted to ...