Name: microfluidic imaging flow cytometry by asymmetric-detection time-stretch optical microscopy (atom)
Uploaded: 2017-06-28T15:00:00
Description: Watch this Scientific Journal Video about Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy ATOM at JoVE.com

感谢杨先生为我们准备MCF-7。这项工作得到了中国香港特别行政区研究资助委员会（项目编号17259316,17207715,17207714,17205215和HKD 720112E），创新与技术支持计划（ITS / 090/14 ）和香港大学发展基金。

样品制备<ol><li> 样品制备（贴壁细胞; MCF-7细胞） <ol><li>从培养箱中取出细胞培养皿，排干培养基。 </li><li>用1x磷酸缓冲盐水（PBS）将培养皿上的细胞冲洗以除去过量的培养基。 </li><li>加入3 mL 0.25％胰蛋白酶溶液至培养皿（直径100 mm），并置于37℃，5％CO 2培养箱中4分钟。 注意：胰蛋白酶溶解细胞的粘合蛋白，使其从培养皿中分离。 </li><li>检查所有的细胞是否使用光学?...

<ol>
	<li>Lau, A. K. S., Wong, T. T. W., Shum, H. C., Wong, K. K. Y., Tsia, K. K., Barteneva, N. S., Vorobjev, I. 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> Imaging Flow Cytometry: Methods and Protocols. , 23-45 (2016).</li><li>Goda, K., Tsia, K. K., Jalali, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serial+time-encoded+amplified+imaging+for+real-time+observation+of+fast+dynamic+phenomena.">Serial time-encoded amplified imaging for real-time observation of fast dynamic phenomena.</a> Nature. 458 (7242), 1145-1149 (2009).</li><li>Goda, K., Jalali, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dispersive+Fourier+transformation+for+fast+continuous+single-shot+measurements.">Dispersive Fourier transformation for fast continuous single-shot measurements.</a> Nat Photon. 7 (2), 102-112 (2013).</li><li>Lau, A. K., Shum, H. C., Wong, K. K., Tsia, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optofluidic+time-stretch+imaging+-+an+emerging+tool+for+high-throughput+imaging+flow+cytometry.">Optofluidic time-stretch imaging - an emerging tool for high-throughput imaging flow cytometry.</a> Lab Chip. 16 (10), 1743-1756 (2016).</li><li>Lau, A. K. S., Tang, A. H. L., Xu, J., Wong, X. W., Y, K. K., Tsia, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+Time+Stretch+for+High-Speed+and+High-Throughput+Imaging+-+From+Single-Cell+to+Tissue-Wide+Scales.">Optical Time Stretch for High-Speed and High-Throughput Imaging - From Single-Cell to Tissue-Wide Scales.</a> IEEE J. Sel. Top. in Quant. Electron. 22 (4), (2016).</li><li>Lei, C., Guo, B., Cheng, Z., Goda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+time-stretch+imaging:+Principles+and+applications.">Optical time-stretch imaging: Principles and applications.</a> Appl Phys Rev. 3 (1), 011102 (2016).</li><li>Lei, C., Guo, B., Cheng, Z., Goda, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+time-stretch+imaging:+Principles+and+applications.">Optical time-stretch imaging: Principles and applications.</a> Applied Physics Reviews. 3 (011102), (2016).</li><li>Goda, K., 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=High-throughput+single-microparticle+imaging+flow+analyzer.">High-throughput single-microparticle imaging flow analyzer.</a> Proc Natl Acad Sci U S A. 109 (29), 11630-11635 (2012).</li><li>Goda, K., Solli, D. R., Tsia, K. K., Jalali, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Theory+of+amplified+dispersive+Fourier+transformation.">Theory of amplified dispersive Fourier transformation.</a> Physical Review A. 80 (4), 043821 (2009).</li><li>Goda, K., 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=High-throughput+single-microparticle+imaging+flow+analyzer.">High-throughput single-microparticle imaging flow analyzer.</a> Proc Natl Acad Sci U S A. 109 (29), 11630-11635 (2012).</li><li>Lai, Q. T. K., 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=High-throughput+time-stretch+imaging+flow+cytometry+for+multi-class+classification+of+phytoplankton.">High-throughput time-stretch imaging flow cytometry for multi-class classification of phytoplankton.</a> Opt. Express. 24 (25), 28170-28184 (2016).</li><li>Chen, C. L., 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=Deep+Learning+in+Label-free+Cell+Classification.">Deep Learning in Label-free Cell Classification.</a> Sci Rep. 6, 21471 (2016).</li><li>Fard, A. 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=Nomarski+serial+time-encoded+amplified+microscopy+for+high-speed+contrast-enhanced+imaging+of+transparent+media.">Nomarski serial time-encoded amplified microscopy for high-speed contrast-enhanced imaging of transparent media.</a> Biomed Opt Express. 2 (12), 3387-3392 (2011).</li><li>Lau, A. K., 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=Interferometric+time-stretch+microscopy+for+ultrafast+quantitative+cellular+and+tissue+imaging+at+1+mum.">Interferometric time-stretch microscopy for ultrafast quantitative cellular and tissue imaging at 1 mum.</a> J Biomed Opt. 19 (7), 76001 (2014).</li><li>Mahjoubfar, A., Chen, C., Niazi, K. R., Rabizadeh, S., Jalali, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Label-free+high-throughput+cell+screening+in+flow.">Label-free high-throughput cell screening in flow.</a> Biomed Opt Express. 4 (9), 1618-1625 (2013).</li><li>Wong, T. 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=Asymmetric-detection+time-stretch+optical+microscopy+(ATOM)+for+ultrafast+high-contrast+cellular+imaging+in+flow.">Asymmetric-detection time-stretch optical microscopy (ATOM) for ultrafast high-contrast cellular imaging in flow.</a> Sci Rep. 4, 3656 (2014).</li><li>Mata, T. M., Martins, A. A., Caetano, N. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microalgae+for+biodiesel+production+and+other+applications:+A+review.">Microalgae for biodiesel production and other applications: A review.</a> Renew Sustainable Energy Rev. 14 (1), 217-232 (2010).</li><li>Irigoien, X., Huisman, J., Harris, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+biodiversity+patterns+of+marine+phytoplankton+and+zooplankton.">Global biodiversity patterns of marine phytoplankton and zooplankton.</a> Nature. 429 (6994), 863-867 (2004).</li><li>Wei, X., 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=Coherent+Laser+Source+for+High+Frame-Rate+Optical+Time-Stretch+Microscopy+at+1.0+&#181;m.">Coherent Laser Source for High Frame-Rate Optical Time-Stretch Microscopy at 1.0 &#181;m.</a> IEEE J. Sel. Top. Quantum Electron. 20 (5), 384-389 (2014).</li><li>Wei, X., 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=Breathing+laser+as+an+inertia-free+swept+source+for+high-quality+ultrafast+optical+bioimaging.">Breathing laser as an inertia-free swept source for high-quality ultrafast optical bioimaging.</a> Opt. Lett. 39 (23), 6593-6596 (2014).</li><li>Xu, Y., Wei, X., Ren, Z., Wong, K. K. Y., Tsia, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrafast+measurements+of+optical+spectral+coherence+by+single-shot+time-stretch+interferometry.">Ultrafast measurements of optical spectral coherence by single-shot time-stretch interferometry.</a> Sci Rep. 6, 27937 (2016).</li><li>Tsia, K. K., Goda, K., Capewell, D., Jalali, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performance+of+serial+time-encoded+amplified+microscope.">Performance of serial time-encoded amplified microscope.</a> Opt. Express. 18 (10), 10016-10028 (2010).</li><li>Alix-Panabi&#232;res, C., Pantel, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Challenges+in+circulating+tumour+cell+research.">Challenges in circulating tumour cell research.</a> Nat Rev Cancer. 14 (9), 623-631 (2014).</li><li>Kling, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beyond+counting+tumor+cells.">Beyond counting tumor cells.</a> Nat Biotech. 30 (7), 578-580 (2012).</li><li>Hodgkinson, C. L., 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=Tumorigenicity+and+genetic+profiling+of+circulating+tumor+cells+in+small-cell+lung+cancer.">Tumorigenicity and genetic profiling of circulating tumor cells in small-cell lung cancer.</a> Nat Med. 20 (8), 897-903 (2014).</li><li>Ozkumur, 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=Inertial+focusing+for+tumor+antigen-dependent+and+-independent+sorting+of+rare+circulating+tumor+cells.">Inertial focusing for tumor antigen-dependent and -independent sorting of rare circulating tumor cells.</a> Sci Transl Med. 5 (179), (2013).</li><li>Hou, H. 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=Isolation+and+retrieval+of+circulating+tumor+cells+using+centrifugal+forces.">Isolation and retrieval of circulating tumor cells using centrifugal forces.</a> Scientific Reports. 3, 1259 (2013).</li><li>Karabacak, N. 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=marker-free+isolation+of+circulating+tumor+cells+from+blood+samples.">marker-free isolation of circulating tumor cells from blood samples.</a> Nat. Protocols. 9 (3), 694-710 (2014).</li><li>Krebs, M. 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=Molecular+analysis+of+circulating+tumour+cells-biology+and+biomarkers.">Molecular analysis of circulating tumour cells-biology and biomarkers.</a> Nat Rev Clin Oncol. 11 (3), 129-144 (2014).</li><li>Wu, J., 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=Ultrafast+Laser-Scanning+Time-Stretch+Imaging+at+Visible+Wavelengths.">Ultrafast Laser-Scanning Time-Stretch Imaging at Visible Wavelengths.</a> Light Sci Appl. , (2016).</li></ol>

在ATOM系统设置过程中需要特别注意的几个技术细节。首先，必须注意的是，不对称/倾斜光谱编码的照明可以在吸收对比度中引入残余相位梯度分量（ 即阴影效应），并影响ATOM中相位梯度对比度的增强。因此，倾斜照明的这种效果应该最小化。第二，应该强调的是，涉及两个副本的时间复用或时间交织方案对成像速度没有显着的影响（ 即，仍然可以维持&gt; MHz的行扫描速率），假定...

光学成像提供了一个强大的工具和基于细胞的测定（几乎）非侵入性地显示许多细胞/亚细胞组分的详细空间分布，从而揭示了细胞的许多形态学，生物物理学和生物分子特征。然而，当必须调查巨大和异质的细胞群体时，从单细胞提取高含量信息的这种能力通常受到损害。这标志着测量吞吐量和内容之间基于细胞的测定中的共同权衡。一个显着的例子是，与传统非成像流式细胞仪相比，添加流式细胞术的成像能力导致吞吐量下降至少1-2个数量级。虽然它可以提供复杂的形态单细胞分析，这是不可能的标准流式细胞仪1 ，成像流式细胞仪通常缺乏足够的吞吐量id以高统计学信度诱导细胞异质性。这对于生物学的新发现和对疾病发病机理的了解是必要的。关键的技术挑战在于通常的光学成像策略固有的速度限制：激光束扫描（ 例如，通过电流镜）和/或图像传感器（ 例如电荷耦合器件（CCD）和互补金属 -氧化物半导体（CMOS））。激光扫描速度本身受到扫描镜的机械惯性的限制，而CCD或CMOS的帧速率受成像速度和灵敏度之间的基本权衡的限制（ 即，增加帧速率导致降低的信号检测灵敏度，反之亦然）。 基于全光学，超快速图像编码机制，光时延成像已经被证明是高通量成像流量cyt的有吸引力的平台米，不需要传统的图像传感器或机械激光扫描2,3 。参考文献4,5,6,7中可以看到时间延伸成像工作原理的详细描述。简而言之，它包括两个可互换的映射步骤：（i）频谱编码（波长空间映射），其中成像的样本的空间坐标映射到光脉冲束8,9的光谱上的不同波长，和（ii）分散傅里叶变换（波长 - 时间映射） 9 ，其中各个激光脉冲的波长分量通过组速度色散（GVD）变换（拉伸）成时间（波长扫描）波形（ 图1 ）。一个重要的时间延伸成像的特征是光学放大，其在抵抗由于超快光检测和GVD损耗引起的灵敏度损失方面起关键作用，从而增强图像信噪比（SNR）而不被光电探测器噪声9污染。由于每个激光脉冲编码与单元的单向流正交的成像样本的线扫描，因此通常超过10MHz的激光重复率来确定有效的线扫描速率。这种超快速操作使得能够以10,000-100,000个细胞/秒的吞吐量（ 即，比传统成像流式细胞术高10-100倍）的无模糊单细胞图像捕获。因此，时间延伸成像可以在高通量，单细胞，基于图像的筛选中找到独特的应用，特别是当需要在相当大的人群中识别未知的异质性或罕见/异常细胞（数千到数百万CEL<html> ls），如罕见癌细胞筛选10或微藻分类11 。 时间延伸成像主要依赖于明场（BF）图像捕获，通过来自细胞2,3,9,10,11的光散射和吸收产生图像对比度。这种无标记的单细胞成像能力可以绕过与荧光标记相关的有害作用，例如细胞毒性和光漂白，并且基于细胞和亚细胞纹理​​和形态为单细胞分析提供有价值的信息。这些无标记参数被证明对于细胞的深层图像分类是有效的，特别是当巨大的细胞群可用时，“外汇公司”&gt; 12。然而，在许多情况下，BF成像不能提供足够的对比度以揭示无标记透明细胞的详细形态。已经开发了不同的无标签，相位对比，时间延伸成像模式，用于在超快帧速率13,14,15下增强成像对比度。在这些技术中，开发了不对称检测时间拉伸光学显微镜（ATOM），以基于类似于Schlieren摄影的概念揭示相位梯度（差分 - 干涉 -​​ 对比 - （DIC）样）的对比度，在超高的微流体速度（高达10 m / s）下，单细胞的免费，高对比度成像16 。通过在光电检测之前部分地阻挡图像编码的光束路径或倾斜光束，可以容易地通过倾斜检测或照明来产生该效果。 ATOM的另一个优点是它的一个能够沿着相反的方向同时获取两个相位梯度对比度。两个对比度图像的强度减法和求和分别从同一行扫描得到差分相位梯度对比度和吸收对比度。这项工作提出了一个详细的协议描述ATOM的实现，包括建立光学设置，样品准备和数据采集和可视化。具体来说，这项工作通过对人类血液细胞，癌细胞和浮游植物（微藻）的单细胞成像证明了ATOM手术。这突出了ATOM对流式细胞术成像的适用性，不仅在生物医学领域，而且在海洋和生物燃料研究17,18 。

<table border="0" cellpadding="0" cellspacing="0" width="789">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:207px;">Nikon Plan Fluorite Physiology Objectives 40X</td>
			<td style="width:71px;">Nikon</td>
			<td style="width:119px;">MRF07420</td>
			<td style="width:393px;">Objective lens (Obj2)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Olympus Plan Fluorite Objective, 0.75 NA, 0.51 mm WD, 40X</td>
			<td style="width:71px;">Olympus</td>
			<td>RMS40X-PF</td>
			<td>Objective lens (Obj1)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:207px;">N-BK7 Plano-Convex Lenses</td>
			<td style="width:71px;">Thorlabs</td>
			<td>LA1145-C</td>
			<td>For relaying spectral shower</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:207px;">FC/APC Fiber Collimation Package</td>
			<td style="width:71px;">Thorlabs</td>
			<td style="width:119px;">F220APC-1064</td>
			<td>For outputing and collecting laser pulses</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:207px;">Pellicle beamsplitter</td>
			<td style="width:71px;">Thorlabs</td>
			<td style="width:119px;">BP145B3</td>
			<td>For making beam replica</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:207px;">Protected silver mirror</td>
			<td style="width:71px;">Thorlabs</td>
			<td style="width:119px;">PF10-03-P01</td>
			<td>For reflecting light</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:207px;">800-1650nm 12GHz single mode DC-coupled NIR Photoreceiver</td>
			<td style="width:71px;">Newport</td>
			<td style="width:119px;">1544-B</td>
			<td>For converting light into electrical signal</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:207px;">Infiniium High-Performance Oscilloscope</td>
			<td style="width:71px;">Agilent</td>
			<td style="width:119px;">DSOX91604A</td>
			<td>To save the light-converted electrical signals</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:207px;">HI1060 optical fiber</td>
			<td style="width:71px;">Corning</td>
			<td style="width:119px;">HI1060</td>
			<td>Optical fiber for time-stretch</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:207px;">YTTERBIUM DOPED FIBER AMPLIFIER</td>
			<td style="width:71px;">Keopsys</td>
			<td style="width:119px;">KPS-STD-BT-YFA-37-BO-SM-111-FA-FA</td>
			<td>Optical in-line amplifier</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:207px;">Holographic grating</td>
			<td style="width:71px;">Wasatch Photonics</td>
			<td style="width:119px;">020305-6</td>
			<td>Grating</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:207px;">Infuse/Withdraw Syringe Pumps</td>
			<td style="width:71px;">Harvard Apparatus</td>
			<td style="width:119px;">PHD 2000</td>
			<td>Syringe pump for sample loading in micro-fluidic channels</td>
		</tr>
	</tbody>
</table>

microfluidic imaging flow cytometry by asymmetric-detection time-stretch optical microscopy (atom)

缩放可测量参数的数量，这是允许进行多维数据分析和因此更高置信度的统计结果，一直是流式细胞术先进发展的主要趋势。值得注意的是，添加高分辨率成像功能允许细胞/亚细胞结构的复杂形态分析。标准流式细胞仪不可能。但是，提高我们的细胞功能知识是有价值的，有利于生命科学研究，临床诊断和环境监测。将成像能力纳入流式细胞术可以降低检测的吞吐量，这主要是由于相机技术中速度和灵敏度的限制。为了克服成像流式细胞术同时保持图像质量面临的这种速度或吞吐量挑战，已经证明了不对称检测时间延伸光学显微镜（ATOM）能够实现高对比度的单细胞图像具有亚细胞分辨率，成像吞吐量高达100,000个细胞/秒。基于通过使用超高速宽带激光脉冲依赖于全光图像编码和检索的常规时间拉伸成像的成像概念，ATOM通过增强未标记/未染色细胞的图像对比来进一步提高成像性能。这是通过访问细胞的相位梯度信息来实现的，其被频谱地编码成单次宽带脉冲。因此，ATOM在单细胞形态和纹理的高通量测量中特别有利 - 指示细胞类型，状态甚至功能的信息。最终，这可能成为用于细胞生物物理表型的强大的成像流式细胞仪平台，补充了当前最先进的基于生物化学标记的细胞测定法。本工作描述了建立ATOM系统关键模块的协议（从光学前端到数据p加工和可视化后端），以及基于ATOM的成像流式细胞术的工作流程，以人类细胞和微藻为例。

<html>这项工作说明了ATOM的两个单细胞成像试验：一个是哺乳动物细胞（人类外周血单个核细胞（PBMC）和乳腺癌细胞（MCF-7）），另一个是浮游植物（Scenedesmus和Chlamydomonas）。第一个实验是由于液体活检对血液中循环肿瘤细胞（CTC）的检测，计数和表征的兴趣日益增加23 。测量从原发肿瘤部位扩散到血液流的CTC的能力允许检测转移性癌症进展...

Watch this Scientific Journal Video about Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy ATOM at JoVE.com

Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy ATOM

该协议描述了用于超快速微流体流中单细胞成像的不对称检测时间拉伸光学显微镜系统的实现及其在成像流式细胞术中的应用。

微流控成像流式细胞仪通过不对称检测时间延伸光学显微镜（ATOM）

Scaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the ...

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Education

JoVE Core

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Electrical Engineering

Engineering

Unit 1

Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM)

Operational Amplifiers

SCIENTISTS IN ACTION

Fabricating Metamaterials Using the Fiber Drawing Method

制造超材料使用光纤拉丝法

Metamaterials are man-made composite materials, fabricated by assembling components much smaller than the wavelength at which they operate 1. They owe ...

科研

工程学

Determining 3D Flow Fields via Multi-camera Light Field Imaging

通过多摄像头的光场成像的三维流场测定

In the field of fluid mechanics, the resolution of computational schemes has outpaced experimental methods and widened the gap between predicted and ...

Optical Trapping of Nanoparticles

纳米粒子的光学捕获

Optical trapping is a technique for immobilizing and manipulating small objects in a gentle way using light, and it has been widely applied in trapping ...

Atom Probe Tomography Studies on the Cu(In,Ga)Se2 Grain Boundaries

Atom Probe Tomography Studies on the CuIn,GaSe2 Grain Boundaries

原子探针层析成像研究在Cu（铟，镓）硒<sub&gt; 2</sub&gt;晶界

Compared with the existent techniques, atom probe tomography is a unique technique able to chemically characterize the internal interfaces at the ...

Analyzing the Movement of the Nauplius 'Artemia salina' by Optical Tracking of Plasmonic Nanoparticles

Analyzing the Movement of the Nauplius 'Artemia salina' by Optical Tracking of Plasmonic Nanoparticles

分析无节幼体的运动“<em&gt;卤虫</em&gt;&#39;由电浆纳米粒子的光学跟踪

We demonstrate how optical tweezers may provide a sensitive tool to analyze the fluidic vibrations generated by the movement of small aquatic organisms. A ...

Coherent anti-Stokes Raman Scattering (CARS) Microscopy Visualizes Pharmaceutical Tablets During Dissolution

Coherent anti-Stokes Raman Scattering CARS Microscopy Visualizes Pharmaceutical Tablets During Dissolution

相干反斯托克斯拉曼散射（CARS）显微可视化药片剂溶出过程中

Traditional pharmaceutical dissolution tests determine the amount of drug dissolved over time by measuring drug content in the dissolution medium. This ...

Writing Bragg Gratings in Multicore Fibers

写布拉格光栅在多核纤维

Fiber Bragg gratings in multicore fibers can be used as compact and robust filters in astronomical and other research and commercial applications. Strong ...

Dynamic Pore-scale Reservoir-condition Imaging of Reaction in Carbonates Using Synchrotron Fast Tomography

用同步快速反应层析成像动态孔隙规模油藏条件下成像碳酸盐

Underground storage permanence is a major concern for carbon capture and storage. Pumping CO2 into carbonate reservoirs has the potential to dissolve ...

Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes

新型光声显微镜与光学相干层析成像在活兔眼中的双模脉络成像

Photoacoustic ocular imaging is an emerging ophthalmic imaging technology that can noninvasively visualize ocular tissue by converting light energy into ...

Generation of Size-controlled Poly (ethylene Glycol) Diacrylate Droplets via Semi-3-Dimensional Flow Focusing Microfluidic Devices

Generation of Size-controlled Poly ethylene Glycol Diacrylate Droplets via Semi-3-Dimensional Flow Focusing Microfluidic Devices

Semi-3-Dimensional 流聚焦微流控装置产生尺寸控制的聚乙二醇二丙烯酸酯液滴

Uniform and size-controllable poly (ethylene glycol) diacrylate (PEGDA) droplets could be produced via the flow focusing process in a microfluidic device. ...