硬件的发展是由美国航空航天局SBIR合同NNX09CA44C和NNX10CA97C支持。对于光学组件和样本装载机演示数据分析是由美国航空航天局的第三阶段合同NNC11CA04C支持。美国航空航天局使用协议IRB＃SA-10-008进行人体血液采集。通过美国国家仪器医疗器械助学计划提供的控制/数据采集软件。模具的微芯片在约翰斯·霍普金斯大学微细加工设备和哈佛大学纳米系统中心作了发言。奥托J.盐水灌装机和卢克贾菲（DNA医学研究所）在2010年夏季美国航空航天局飞行视频的工作人员在飞行过程中每周提供录像辅助机架装配。卡洛斯·巴里恩托斯（DNA医学研究所）提供的照片和图形的援助。特别感谢协助获得空间环境对技术2010计划，美国航天局的低重力办公室，人力适应与对策部，国家航空航天局格伦研究中心，ZIN技术，和人类的研究计划。

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尤金Y.陈康迪斯裴和朱莉娅Z.夏普的发明人对通过DNA医学研究所，一个商业实体，申请相关技术的专利。

这里所描述的方法，使主要技术组件（装样，微混合和光学检测）的有效证明在2010年快速抛物线飞行，用比较的结果，以地面试验。这里所描述的培训和SOP方法特别有效，并帮助照亮依靠实践示威不会板载了抛物线飞行工具等“拐杖”的存在。 需要改进的地方，包括遏制和布局。定制亚克力成分可能不是遏制的目的足够强大。在“手套”框被重力过渡期间遭飞行的乘客，并?...

目前的空间准备健康诊断的不足之处提出了一个限​​制因素，以更深入的载人太空探索。诊断需要是全面的，易于在低重力来使用，并且相对地不受发射和航天（ 例如 ，高g力，振动，辐射，温度变化，和机舱压力的变化）的压力。在护理点的检测（POCT）的发展可以通过使用更小的患者的标本（ 例如 ，一个手指刺），更简单和更小的流体学（ 即 ，微流体）转化为有效的航天溶液，并降低电功率的要求，除其他优势。 流式细胞术是有吸引力的方法为在空间的POC，因为该技术的广泛的用途，包括对细胞计数和生物标志物的定量分析，以及显著小型化的潜力。上一页空间相关的流式细胞仪，包括“核包装efficiency&#39;（NPE）的工具，用于同时进行电弧灯诱导荧光和电子音量（库尔特体积）测量1-4中 ，一个相对较小的台式流式细胞仪代表“第一代流式细胞仪中的零重力&#39;5数据的实时流，一个“无鞘微仪”能够4-和5-部分白血细胞（WBC）分类计数用预处理5微升全血样品6-9，以及“光纤为基础的”流式细胞仪最近在国际板载测试空间站10。 通常进行板载了使用近似抛物线飞行弹道模拟失重（ 例如 ，零重力，火星的重力）11的选择水平降低重力飞机的潜在空间应用评估诊断技术。评估是具有挑战性的，因为飞行的机会是有限的，repet微重力可持续的竞争短窗可以使它很难评估方法或通常需要不间断的时间超过20-40秒长的过程，并演示可能需要飞行12-15不要轻易使用额外的设备。此外，在使用或设计，减少了重力作用的体外诊断（IVD）技术先前的示威是有限的，大量的工作仍然未公布。除了 ​​上述的流式细胞仪，在文献中描述的其他空间相关的IVD-技术包括全血染色装置，用于免疫的应用程序16，一个自动化的基于照相机的流式细胞仪12，手持式的临床分析仪，用于集成电势，电流分析法，和电导12,17，一个微流体“T形感应器”装置，用于分析物的定量分析，依赖于扩散为基础的混合和分离18，和一个旋转的&#39;实验室上的CD“诊断平台19,20。初来乍到低重力测试也可能看起来不相关的体外诊断抛物线飞行表演时，试图使设备评估可能的（或搞清楚什么是可能的）。从具有证据充分的飞行准备，飞行中的战略，以及飞行测试设备等以前的医学或生物实验演示包括在表1中 15，21-35。这些可以是信息性由于包含的手册中飞行任务，使用专门的设备，并且实验遏制。 <table border="0" cellpadding="0" cellspacing="0"><tbody><tr height="17"><td height="17" style="height:17px;width:133px;"> 类别 </td><td style="width:497px;"> 示例 </td></tr><tr height="40"><td height="40" style="height:40px;width:133px;">紧急医疗</td><td style="width:497px;">气管插管（喉镜引导下，对马尼克在）21，心脏生命支持（麻醉猪）22 </td></tr><tr height="20"><td height="20" style="height:20px;width:133px;">外科护理</td><td style="width:497px;">腹腔镜手术（视频模拟23，对麻醉的猪24,25） </td></tr><tr height="41"><td height="41" style="height:41px;width:133px;">医学影像学或生理学评估</td><td style="width:497px;">超声与下体负压室26，多普勒流量计（头戴式）27，中心静脉压监测器28 </td></tr><tr height="65"><td height="65" style="height:65px;width:133px;">专业的生物设备</td><td style="width:497px;">酶标仪（和飞行中的手套箱中）29，对细胞周期的实验30，显微镜温度控制系统（明场，相位相反，和多通道荧光能力）15，毛细管电泳设备连接到视频显微镜31 </td></tr><tr height="20"><td height="20" style="height:20px;width:133px;">其他</td><td style="width:497px;">植物收割用钳子32，只包含33,34和35鱼类观察</td></tr></tbody></table> 表1.抛物线飞行有很好的描述方法/试验示范的例子 为了扩大在前面的例子中，提供更深入地了解成功的飞行演示中，我们提出了模块化和适应性强的程序结构的原型与相关的微混技术的流式细胞仪为抛物线飞行试验台的组成部分和操作。该钻机能装样，微混合和荧光粒子探测的演示，并进行了测试机上，2010年美国航空航天局方便获取的空间环境（FAST）抛物线fligHTS，飞行距离9月29日至十月一日，2010年这些示威活动从一开始，中间和结束拉，分别在指尖大小的血样被载入，稀释或者与试剂混合的潜在设备的工作流程，并通过光学分析检测。流式细胞仪缩放流入一个紧凑的单元需要创新和精心挑选的一部分。定制和现成的，现成的组件是用在这里，所选择的最后一个组件的选择最好的早期近似的，以及可能适用于其他创新的设计。下面的原型组件选择一个大纲，设置是在作为骨架钻机装配支撑结构描述。原型部件被分配的地点，固定，并伴有必要的成功实验的附加组件。注意然后转移到涉及标准操作程序（SOP）的开发，培训，物流等较为抽象的过程。最后，演示，具体程序描述。此处所描述的策略和支撑钻机部件（ 例如 ，显微镜，丙烯酸箱等 ）的选择，尽管执行此处为特定原型，说话相关测试任何血液诊断设备中减小的重力环境的一般问题和挑战。 在航班2010年2月球重力（达到约1/6地球重力）和两个微重力航班原定跨越4天，虽然最终这些被跨3天改期。示威板载了修改私人经营，窄体喷气客机36进行。每个航班提供30-40抛物线，每产生约20秒的引力（大致1.8克），随后由20-25秒的减少重力条件下。被处决的一半抛物线后，飞机停了一段平飞约5-10分钟，以使飞机调头后脑勺朝着陆点，而PErforming的抛物线的其余部分。

<table border="0" cellpadding="0" cellspacing="0" width="1933">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Name of Material/ Equipment</td>
			<td style="width:287px;">Company</td>
			<td style="width:135px;">Catalog Number</td>
			<td style="width:1143px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Micro air pump</td>
			<td style="width:287px;">Smart Products, Inc.</td>
			<td style="width:135px;">AP-2P02A</td>
			<td>Max pressure = 6.76 psi; 1.301&#8221; x 0.394&#8221; x 0.650&#8221; , 0.28 oz (8 g); available direct from Smart Products</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Differential pressure sensor</td>
			<td style="width:287px;">Honeywell International, Inc.</td>
			<td style="width:135px;">ASDX015D44R</td>
			<td>Range&#160; of&#160; 0-15psi; 0.974&#34; x 0.550&#34; x 0.440&#34;, 0.09 oz (2.565 g); suppliers include Digi-Key and Mouser Electronics</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Rigid plastic vial (small size)</td>
			<td style="width:287px;">Loritz &#38; Associates, Inc.</td>
			<td style="width:135px;">55-05</td>
			<td>Polystyrene; ID 0.81&#34; (20.6 mm), IH 2.06&#34; (52.4 mm); available direct from LA Container Inc.; similar product available from Dynalab Corp.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Rigid plastic vial (larger size)</td>
			<td style="width:287px;">Loritz &#38; Associates, Inc.</td>
			<td style="width:135px;">55-140</td>
			<td>Polystyrene; ID 1.88&#34; (47.6 mm), IH 3.31&#34; (84.1 mm); available direct from LA Container Inc.; similar product available from Dynalab Corp.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">latex examination gloves</td>
			<td style="width:287px;">dynarex corporation</td>
			<td style="width:135px;">2337</td>
			<td>Middle finger used for latex diaphragm in fluid source vial.&#160; Other brands (e.g., Aurelia &#174;&#160; Vibrant &#8482;) acceptable.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Optical glue</td>
			<td style="width:287px;">Norland Products</td>
			<td style="width:135px;">NOA 88</td>
			<td>Low outgassing adhesive; available direct from Norland; Also available from Edmund Optics Inc.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">3-way solenoid valves</td>
			<td style="width:287px;">The LEE Company</td>
			<td style="width:135px;">LHDA0531115H</td>
			<td>Gas valves, but can function with liquid; 1.29 &#34; L, 0.28 &#34; D.&#160; Discontinued product.&#160; Similar products available from The LEE Company.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Volumetric water flowmeter</td>
			<td style="width:287px;">OMEGA Engineering inc.&#160;</td>
			<td style="width:135px;">FLR-1602A</td>
			<td>Non-contacting flow rate meter strongly preferred.&#160; We recommend SENSIRION LG16 OEM Liquid Flow Sensor for flow rates from nl/min up to 5 ml/min.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">PCD-mini photon detector&#160;</td>
			<td style="width:287px;">Sensl</td>
			<td style="width:135px;">PCDMini-00100</td>
			<td style="width:1143px;">For fluorescence detection; available direct from Sensl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Accelerometer</td>
			<td style="width:287px;">Crossbow Technology, Inc.</td>
			<td style="width:135px;">CXL02LF3</td>
			<td>3-demensional force detection.&#160; Supplied to DMI by NASA.&#160; Similar product available from Vernier Software &#38; Technology, LLC.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Stereomicroscope</td>
			<td style="width:287px;">AmScope</td>
			<td style="width:135px;">SE305R-AZ-E</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">CCD Camera</td>
			<td style="width:287px;">Thorlabs</td>
			<td style="width:135px;">DCU223C</td>
			<td>1024 x 768 Resolution, Color, USB 2.0; available direct from Thorlabs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">USB and Trigger Cable (In/Out) for CCD Camera</td>
			<td style="width:287px;">Thorlabs</td>
			<td style="width:135px;">CAB-DCU-T1</td>
			<td>Available direct from Thorlabs</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Microbore tubing</td>
			<td style="width:287px;">Saint-Gobain Corporation</td>
			<td>AAD04103</td>
			<td>Tygon&#174;; ID 0.02&#34;, OD 0.06&#34;, 500ft, 0.02&#34; wall. Suppliers: VWR, Thermo Fisher Scientific Inc.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Hollow steel pins</td>
			<td style="width:287px;">New England Small Tube</td>
			<td style="width:135px;">(Custom)</td>
			<td>&#160;0.025&#34; OD, 0.017&#34; ID, 0.500&#8221; L, stainless steel tube, type 304, cut, deburred, passivated; enable microbore tubing connections, chip tubing connections</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Slide clamp</td>
			<td style="width:287px;">World Precision Instruments, Inc.</td>
			<td style="width:135px;">14042</td>
			<td>Available direct from World Precision Instruments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Leur adaptor pieces</td>
			<td style="width:287px;">World Precision Instruments, Inc.</td>
			<td style="width:135px;">14011</td>
			<td>Available direct from World Precision Instruments</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Silicon wafer</td>
			<td style="width:287px;">Addison Engineering, Inc.</td>
			<td style="width:135px;"></td>
			<td>6&#34; diameter; for SU-8 mold fabrication</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;width:368px;">Polydimethylsiloxane (PDMS) elastomer base</td>
			<td style="width:287px;">Dow Corning</td>
			<td style="width:135px;">3097366-1004</td>
			<td>Supplier: Global Industrial SLP, LLC</td>
		</tr>
		<tr height="18">
			<td height="18" style="height:19px;width:368px;">Polydimethylsiloxane (PDMS) elastomer curing agent</td>
			<td style="width:287px;">Dow Corning</td>
			<td style="width:135px;">3097358-1004</td>
			<td>Supplier: Global Industrial SLP, LLC</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Needle (23 gauge), bevel tip</td>
			<td style="width:287px;">Terumo Medical Corporation</td>
			<td style="width:135px;">NN-2338R</td>
			<td>Ultra thin wall; 23G x 1.5&#34;; 22G also usable; suppliers: Careforde, Inc.,&#160; Port City Medical</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Dispensing needle (23 gauge), blunt tip</td>
			<td style="width:287px;">CML Supply</td>
			<td style="width:135px;">901-23-100</td>
			<td>23Gx 1&#34;;&#160; available from CML Supply</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Rotary tool</td>
			<td style="width:287px;">Robert Bosch Tool Corporation</td>
			<td style="width:135px;">1100-01</td>
			<td>Dremel&#174; 1100-01 Stylus&#8482;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Cover glass</td>
			<td style="width:287px;">Thermo Fisher Scientific, Inc.</td>
			<td style="width:135px;">12-518-105E</td>
			<td>Gold Seal&#8482; noncorrosive borosilicate glass; for PDMS chip cover; 24x60 mm; available from Thermo Fisher Scientific, Inc.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Vacuum pump</td>
			<td style="width:287px;">Mountain</td>
			<td style="width:135px;">MTN8407</td>
			<td>For degassing PDMS; supplier:&#160; Ryder System, Inc.&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Vacuum chamber</td>
			<td style="width:287px;">Thermo Fisher Scientific, Inc.</td>
			<td style="width:135px;">5311-0250</td>
			<td>Nalgene&#8482; Transparent Polycarbonate; available from Thermo Fisher Scientific, Inc.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Plasma cleaner</td>
			<td style="width:287px;">Harrick Plasma</td>
			<td style="width:135px;">PDC-32G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Hand magnifier</td>
			<td style="width:287px;">Mitutoyo</td>
			<td style="width:135px;">183-131</td>
			<td>Use in reverse direction to enable viewing at ~15&#34;.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Ethanol</td>
			<td style="width:287px;">CAROLINA</td>
			<td style="width:135px;">861283</td>
			<td>For chip cleaning. Dilute to 70% using millipore water.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Water purification system</td>
			<td style="width:287px;">Thermo Fisher Scientific, Inc.</td>
			<td style="width:135px;">D11901</td>
			<td>Available direct from Thermo Fisher Scientific, Inc.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Optomechanical translation mounts</td>
			<td style="width:287px;">Thorlabs</td>
			<td style="width:135px;">K6X</td>
			<td>6-Axis Kinematic Optic Mount; discontinued product; new product (K6XS) available direct from Thorlabs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Laptop</td>
			<td style="width:287px;">Hewlett-Packard</td>
			<td style="width:135px;">VP209AV</td>
			<td>HP Pavilion Laptop running Windows 7</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Laptop tray (spring loaded)</td>
			<td style="width:287px;">National Products, INC.</td>
			<td style="width:135px;">RAM-234-3&#160;</td>
			<td>RAM Tough-Tray&#8482;. Can accommodate 10 to 16 inch wide laptops.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">USB splitter</td>
			<td style="width:287px;">Connectland Technology Limited</td>
			<td style="width:135px;">3401167</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:18px;width:368px;">USB Data Acquisition Cards (8 analog input, 12 digital I/O)</td>
			<td style="width:287px;">National Instruments</td>
			<td style="width:135px;">NI USB-6008</td>
			<td>12-Bit, 10 kS/s Low-Cost Multifunction DAQ</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:368px;">USB Data Acquisition Cards (16 analog input, 32 digital I/O)</td>
			<td style="width:287px;">National Instruments</td>
			<td style="width:135px;">NI USB-6216</td>
			<td>16-Bit, 400 kS/s Isolated M Series MIO DAQ, Bus-Powered</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Control/acquisition Software</td>
			<td style="width:287px;">National Instruments</td>
			<td style="width:135px;">LabVIEW 2009</td>
			<td>Custom coded National Instruments (NI) LabVIEW&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:22px;width:368px;">3D Solid Modeling Software</td>
			<td style="width:287px;">Dassault Syst&#232;mes SolidWorks Corp.</td>
			<td style="width:135px;">SolidWorks 2011</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">2D Modeling Software</td>
			<td style="width:287px;">AUTODESK</td>
			<td style="width:135px;">AutoCAD LT 2008</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Vertical equipment rack</td>
			<td style="width:287px;">(NASA provided)</td>
			<td style="width:135px;">N/A</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:368px;">Solid aluminum optical breadboard</td>
			<td style="width:287px;">Thorlabs</td>
			<td style="width:135px;">MB2424</td>
			<td>24&#34; x 24&#34; x 1/2&#34;, 1/4&#34;-20 Taps; available direct from Thorlabs</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Industrial grade steel and hardener</td>
			<td style="width:287px;">The J-B Weld Company</td>
			<td style="width:135px;"></td>
			<td>J-B Weld Steel Reinforced Epoxy Glue</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Micro-hematocrit capillary&#160;</td>
			<td style="width:287px;">Fisher Scientific</td>
			<td style="width:135px;">22-362-574</td>
			<td>inner diamter 1.1 to 1.2 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">1 mL syringes</td>
			<td style="width:287px;">Henke-Sass, Wolf</td>
			<td>4010.200V0</td>
			<td>NORM-JECT&#174;; supplier: Grainger, Inc.</td>
		</tr>
		<tr height="80">
			<td height="80" style="height:81px;width:368px;">Human red blood cells</td>
			<td style="width:287px;">Innovative Research</td>
			<td>IPLA-WB3</td>
			<td style="width:1143px;">Tested and found negative by supplier for: HBsAg, HCV, HIV-1, HIV-2, HIV-1Ag or HIV 1-NAT, ALT, and syphilis by FDA-Approved Methods.&#160; Because no test methods can guarantee with 100% certainty the absence of an infectious agent, human derived products should be handled as suggested in the U.S. Department of Health and Human Services Manual on BIOSAFETY IN MICROBIOLOGICAL AND BIOMEDICAL LABORATORIES, FOR POTENTIALLY INFECTIOUS HUMAN SERUM OR BLOOD SPECIMENS</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Phosphate buffered saline concentrate</td>
			<td>P5493</td>
			<td style="width:135px;">SIGMA</td>
			<td>10x; diluted to 1x</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tween</td>
			<td>P9416</td>
			<td style="width:135px;">SIGMA</td>
			<td>TWEEN&#174; 20</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Centrifuge</td>
			<td style="width:287px;">LW Scientific</td>
			<td style="width:135px;">STRAIGHT8-5K</td>
			<td>Swing-Out 8-place Centrifuge.&#160; Available through authorized dealers.&#160; Other centrifuges available direct from LW Scientific.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">HD video recorder</td>
			<td style="width:287px;">Sony</td>
			<td style="width:135px;">MHS-CM5</td>
			<td></td>
		</tr>
		<tr height="52">
			<td height="52" style="height:53px;width:368px;">Orange fluorescent nucleic acid stain</td>
			<td style="width:287px;">Invitrogen</td>
			<td>S-11364</td>
			<td style="width:1143px;">SYTO&#174; 83 Orange Fluorescent Nucleic Acid Stain.&#160; Stored in DMSO solvent. Always wear reccommended Personal Protective Equipment. No special handling 
			advice required.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:368px;">Fluorescent counting beads</td>
			<td style="width:287px;">Invitrogen</td>
			<td>MP 36950</td>
			<td>CountBright&#8482; Absolute Counting Beads.&#160; Always wear reccommended Personal Protective Equipment. No special handling advice required.</td>
		</tr>
	</tbody>
</table>

reduced-gravity environment hardware demonstrations of a prototype miniaturized flow cytometer and companion microfluidic mixing technology

直到最近，收集宇航员的血液样本中飞行，运到陆地上的航天飞机，并在地面的实验室进行分析。如果人类前往超越低地球轨道，走向太空就绪，点的护理的过渡（POC）测试是必需的。这样的测试必须是全面的，容易降低重力环境下进行，并且不会受到发射和航天的压力。无数POC设备已经被开发，以模仿实验室规模的对应，但最有窄的应用和一些具有在飞行中，减少的重力环境可证实的用途。事实上，在降低重心的生物医学诊断的示威完全限制，使得组件的选择和一些后勤方面的挑战很难寻求测试新技术的时候进场。为了帮助填补这一空白，我们提出了模块化方法的原型血液诊断设备的建设和运营及其相关的P抛物飞行试验台符合标准的飞行试验机载抛物线飞行，减少了重力飞机。该方法首先侧重于钻机装配在飞机上，减少了重力流式细胞仪的测试和同伴微流控芯片的混合。组件是适合于其他的设计和一些定制组件，比如一个微量样品装载器和所述微混合器可以是特别感兴趣的。然后该方法将焦点转移到飞行准备，通过提供指导和建议，以一个成功的飞行试验方面用户培训，标准作业程序（SOP）的发展等问题做准备。最后，在飞行中特定于我们的示范实验步骤进行了描述。

对于微混合器示范代表性的结果显示在图7中 ，作为观看由安装在立体显微镜CCD照相机。混合可以在视觉上评价在沿着螺旋的任何点，以及在涉及两组流体实验的出口通道：血液/盐水和蓝色/黄色染料。该二维图像的定量分析可以包括确定在整个信道宽度遮阳均匀性在不同的区域，如图中的其他出版物38-40。见补充图1进一步的细节。见补充图2演示泡处...

Watch this Scientific Journal Video about Reduced-gravity Environment Hardware Demonstrations of a Prototype Miniaturized Flow Cytometer and Companion Microfluidic Mixing Technology at JoVE.com

Reduced-gravity Environment Hardware Demonstrations of a Prototype Miniaturized Flow Cytometer and Companion Microfluidic Mixing Technology

航天血液诊断需要创新。少数示威已经公布出在飞行中，减少了重力医疗诊断技术。这里我们提出了结构和操作一个抛物线飞行试验台上对护理点的一个原型流式细胞仪的设计，与组分和制备策略适应其它设置方法。

样机小型化流式细胞仪及配套的混合微流控技术降低重力环境硬件演示

Until recently, astronaut blood samples were collected in-flight, transported to earth on the Space Shuttle, and analyzed in terrestrial laboratories. If ...

reduced-gravity-environment-hardware-demonstrations-prototype

Research

JoVE Journal

Biology

13.6K 次观看.  DNA Medicine Institute. 航天血液诊断需要创新。少数示威已经公布出在飞行中，减少了重力医疗诊断技术。这里我们提出了结构和操作一个抛物线飞行试验台上对护理点的一个原型流式细胞仪的设计，与组分和制备策略适应其它设置方法。 

视频: Reduced-gravity Environment Hardware Demonstrations of a Prototype Miniaturized Flow Cytometer and Companion Microfluidic Mixing Technology

A Microfluidic Device with Groove Patterns for Studying Cellular Behavior

We describe a microfluidic device with microgrooved patterns for studying cellular behavior. This microfluidic platform consists of a top fluidic channel and a bottom microgrooved substrate. To fabricate the microgrooved channels, a top poly(dimethylsiloxane) (PDMS) mold containing the impression of the microfluidic channels was aligned and bonded to a microgrooved substrate. Using this device, mouse fibroblast cells were immobilized and patterned within microgrooved substrates (25, 50, 75, and 100 &mu;m wide). To study apoptosis in a microfluidic device, media containing hydrogen peroxide, Annexin V, and propidium iodide was perfused into the fluidic channel for 2 hours. We found that cells exposed to the oxidative stress became apoptotic. These apoptotic cells were confirmed by Annexin V that bound to phosphatidylserine at the outer leaflet of the plasma membrane during the apoptosis process. Using this microfluidic device with microgrooved patterns, the apoptosis process was observed in real-time and analyzed by using an inverted microscope containing an incubation chamber (37&deg;C, 5% CO2). Therefore, this microfluidic device incorporated with microgrooved substrates could be useful for studying the cellular behavior and performing high-throughput drug screening.

We describe a protocol for the fabrication of microfluidic devices that can enable cell capture and culture. In this approach patterned microstructures such as grooves within microfluidic channels are used to create low shear stress regions within which cell can dock.

微流体装置与Groove的模式，为研究细胞行为

We describe a microfluidic device with microgrooved patterns for studying cellular behavior.  This microfluidic platform consists of a top fluidic channel ...

科研

生物学

A Gradient-generating Microfluidic Device for Cell Biology

 The fabrication and operation of a gradient-generating microfluidic device for studying cellular behavior is described.  A microfluidic platform is an enabling experimental tool, because it can precisely manipulate fluid flows, enable high-throughput experiments, and generate stable soluble concentration gradients.  Compared to conventional gradient generators, poly(dimethylsiloxane) (PDMS)-based  microfluidic devices can generate stable concentration gradients of growth factors with well-defined profiles.  Here, we developed simple gradient-generating microfluidic devices with three separate inlets.  Three microchannels combined into one microchannel to generate concentration gradients.  The stability and shape of growth factor gradients were confirmed by fluorescein isothyiocyanate (FITC)-dextran with a molecular weight similar to epidermal growth factor (EGF).  Using this microfluidic device, we demonstrated that fibroblasts exposed to concentration gradients of EGF migrated toward higher concentrations.  The directional orientation of cell migration and motility of migrating cells were quantitatively assessed by cell tracking analysis.  Thus, this gradient-generating microfluidic device might be useful for studying and analyzing the behavior of migrating cells.

We describe a protocol for the microfabrication of the gradient-generating microfluidic device that can generate spatial and temporal gradients in well-defined microenvironment. In this approach, the gradient-generating microfluidic device can be used to study directed cell migration, embryogenesis, wound healing, and cancer metastasis.

一个渐变产生的微流体装置，细胞生物学

 The fabrication and operation of a gradient-generating microfluidic device for studying cellular behavior is described.  A microfluidic platform is an ...

CD4+ T-Lymphocyte Capture Using a Disposable Microfluidic Chip for HIV

CD4 + T淋巴细胞艾滋病毒的捕获使用一次性微流体芯片

A Multi-compartment CNS Neuron-glia Co-culture Microfluidic Platform

We present a novel multi-compartment neuron co-culture microsystem platform for in vitro CNS axon-glia interaction research, capable of conducting up to six independent experiments in parallel for higher-throughput. We developed a new fabrication method to create microfluidic devices having both micro and macro scale structures within the same device through a single soft-lithography process, enabling mass fabrication with good repeatability.
The multi-compartment microfluidic co-culture platform is composed of one soma compartment for neurons and six axon/glia compartments for oligodendrocytes (OLs). The soma compartment and axon/glia compartments are connected by arrays of axon-guiding microchannels that function as physical barriers to confine neuronal soma in the soma compartment, while allowing axons to grow into axon/glia compartments. OLs loaded into axon/glia compartments can interact only with axons but not with neuronal soma or dendrites, enabling localized axon-glia interaction studies. The microchannels also enabled fluidic isolation between compartments, allowing six independent experiments to be conducted on a single device for higher throughput.
Soft-lithography using poly(dimethylsiloxane) (PDMS) is a commonly used technique in biomedical microdevices. Reservoirs on these devices are commonly defined by manual punching. Although simple, poor alignment and time consuming nature of the process makes this process not suitable when large numbers of reservoirs have to be repeatedly created. The newly developed method did not require manual punching of reservoirs, overcoming such limitations. First, seven reservoirs (depth: 3.5 mm) were made on a poly(methyl methacrylate) (PMMA) block using a micro-milling machine. Then, arrays of ridge microstructures, fabricated on a glass substrate, were hot-embossed against the PMMA block to define microchannels that connect the soma and axon/glia compartments. This process resulted in macro-scale reservoirs (3.5 mm) and micro-scale channels (2.5 &mu;m) to coincide within a single PMMA master. A PDMS replica that served as a mold master was obtained using soft-lithography and the final PDMS device was replicated from this master.
Primary neurons from E16-18 rats were loaded to the soma compartment and cultured for two weeks. After one week of cell culture, axons crossed microchannels and formed axonal only network layer inside axon/glia compartments. Axons grew uniformly throughout six axon/glia compartments and OLs from P1-2 rats were added to axon/glia compartments at 14 days in vitro for co-culture.

We developed a novel multi-compartment neuron co-culture microsystem platform for in vitro CNS axon-glia interaction research. The platform is capable of conducting up to six independent experiments in parallel and was fabricated using a newly developed macro/micro hybrid fabrication method.

一个多室中枢神经系统的神经胶质细胞共培养的微流体平台

We present a novel multi-compartment neuron co-culture microsystem platform for in vitro CNS axon-glia interaction research, capable of conducting up to ...

The Microfluidic Probe: Operation and Use for Localized Surface Processing

Microfluidic devices allow assays to be performed using minute amounts of sample and have recently been used to control the microenvironment of cells. Microfluidics is commonly associated with closed microchannels which limit their use to samples that can be introduced, and cultured in the case of cells, within a confined volume. On the other hand, micropipetting system have been used to locally perfuse cells and surfaces, notably using push-pull setups where one pipette acts as source and the other one as sink, but the confinement of the flow is difficult in three dimensions. Furthermore, pipettes are fragile and difficult to position and hence are used in static configuration only. 
The microfluidic probe (MFP) circumvents the constraints imposed by the construction of closed microfluidic channels and instead of enclosing the sample into the microfluidic system, the microfluidic flow can be directly delivered onto the sample, and scanned across the sample, using the MFP. . The injection and aspiration openings are located within a few tens of micrometers of one another so that a microjet injected into the gap is confined by the hydrodynamic forces of the surrounding liquid and entirely aspirated back into the other opening. The microjet can be flushed across the substrate surface and provides a precise tool for localized deposition/delivery of reagents which can be used over large areas by scanning the probe across the surface. 
In this video we present the microfluidic probe1 (MFP). We explain in detail how to assemble the MFP, mount it atop an inverted microscope, and align it relative to the substrate surface, and finally show how to use it to process a substrate surface immersed in a buffer.

In this video we present the microfluidic probe1 (MFP). We explain in detail how to assemble the MFP, mount it atop an inverted microscope, and align it relative to the substrate surface, and finally show how to use it to process a substrate surface immersed in a buffer solution.

微探针：操作和使用的局部表面处理

Microfluidic devices allow assays to be performed using minute amounts of sample and have recently been used to control the microenvironment of cells. ...

Platelet Adhesion and Aggregation Under Flow using Microfluidic Flow Cells

Platelet aggregation occurs in response to vascular injury where the extracellular matrix below the endothelium has been exposed. The platelet adhesion cascade takes place in the presence of shear flow, a factor not accounted for in conventional (static) well-plate assays. This article reports on a platelet-aggregation assay utilizing a microfluidic well-plate format to emulate physiological shear flow conditions. Extracellular proteins, collagen I or von Willebrand factor are deposited within the microfluidic channel using active perfusion with a pneumatic pump. The matrix proteins are then washed with buffer and blocked to prepare the microfluidic channel for platelet interactions. Whole blood labeled with fluorescent dye is perfused through the channel at various flow rates in order to achieve platelet activation and aggregation. Inhibitors of platelet aggregation can be added prior to the flow cell experiment to generate IC50 dose response data.

The platelet adhesion cascade takes place in the presence of shear flow, a factor not accounted for in conventional (static) well-plate assays. This article reports on a platelet-aggregation assay utilizing a microfluidic well-plate format to emulate physiological shear flow conditions.

根据流量的血小板粘附和聚集，使用微流控流细胞

Platelet aggregation occurs in response to vascular injury where the extracellular matrix below the endothelium has been exposed. The platelet adhesion ...

Microfluidic Co-culture of Epithelial Cells and Bacteria for Investigating Soluble Signal-mediated Interactions

The human gastrointestinal (GI) tract is a unique environment in which intestinal epithelial cells and non-pathogenic (commensal) bacteria coexist. It has been proposed that the microenvironment that the pathogen encounters in the commensal layer is important in determining the extent of colonization. Current culture methods for investigating pathogen colonization are not well suited for investigating this hypothesis as they do not enable co-culture of bacteria and epithelial cells in a manner that mimics the GI tract microenvironment. Here we describe a microfluidic co-culture model that enables independent culture of eukaryotic cells and bacteria, and testing the effect of the commensal microenvironment on pathogen colonization. The co-culture model is demonstrated by developing a commensal Escherichia coli biofilm among HeLa cells, followed by introduction of enterohemorrhagic E. coli (EHEC) into the commensal island, in a sequence that mimics the sequence of events in GI tract infection.

This protocol describes a microfluidic co-culture model for simultaneous and localized culture of epithelial cells and bacteria. This model can be used for investigating the role of different soluble molecular signals on pathogenesis as well as screen the effectiveness of putative probiotic bacterial strains.

微流体调查可溶性信号介导的相互作用的上皮细胞和细菌的共同文化

The human gastrointestinal (GI) tract is a unique environment in which intestinal epithelial cells and non-pathogenic (commensal) bacteria coexist. It has ...

Quantifying Mixing using Magnetic Resonance Imaging

Mixing is a unit operation that combines two or more components into a homogeneous mixture. This work involves mixing two viscous liquid streams using an in-line static mixer. The mixer is a split-and-recombine design that employs shear and extensional flow to increase the interfacial contact between the components. A prototype split-and-recombine (SAR) mixer was constructed by aligning a series of thin laser-cut Poly (methyl methacrylate) (PMMA) plates held in place in a PVC pipe. Mixing in this device is illustrated in the photograph in Fig. 1. Red dye was added to a portion of the test fluid and used as the minor component being mixed into the major (undyed) component. At the inlet of the mixer, the injected layer of tracer fluid is split into two layers as it flows through the mixing section. On each subsequent mixing section, the number of horizontal layers is duplicated. Ultimately, the single stream of dye is uniformly dispersed throughout the cross section of the device.
 Using a non-Newtonian test fluid of 0.2% Carbopol and a doped tracer fluid of similar composition, mixing in the unit is visualized using magnetic resonance imaging (MRI). MRI is a very powerful experimental probe of molecular chemical and physical environment as well as sample structure on the length scales from microns to centimeters. This sensitivity has resulted in broad application of these techniques to characterize physical, chemical and/or biological properties of materials ranging from humans to foods to porous media 1, 2. The equipment and conditions used here are suitable for imaging liquids containing substantial amounts of NMR mobile 1H such as ordinary water and organic liquids including oils. Traditionally MRI has utilized super conducting magnets which are not suitable for industrial environments and not portable within a laboratory (Fig. 2). Recent advances in magnet technology have permitted the construction of large volume industrially compatible magnets suitable for imaging process flows. Here, MRI provides spatially resolved component concentrations at different axial locations during the mixing process. This work documents real-time mixing of highly viscous fluids via distributive mixing with an application to personal care products.

Magnetic resonance imaging (MRI) provides a powerful tool to evaluate the effectiveness of process equipment during operation. We discuss the use of MRI to visualize mixing in a static mixer. The application is relevant to personal care products, but can be applied to a broad range of food, chemical, biomass and biological fluids.

量化混合使用磁共振成像

Mixing is a unit operation that combines two or more components into a homogeneous mixture.  This work involves mixing two viscous liquid streams using an ...

Chromosomics: Detection of Numerical and Structural Alterations in All 24 Human Chromosomes Simultaneously Using a Novel OctoChrome FISH Assay

Fluorescence in situ hybridization (FISH) is a technique that allows specific DNA sequences to be detected on metaphase or interphase chromosomes in cell nuclei1. The technique uses DNA probes with unique sequences that hybridize to whole chromosomes or specific chromosomal regions, and serves as a powerful adjunct to classic cytogenetics. For instance, many earlier studies reported the frequent detection of increased chromosome aberrations in leukemia patients related with benzene exposure, benzene-poisoning patients, and healthy workers exposed to benzene, using classic cytogenetic analysis2. Using FISH, leukemia-specific chromosomal alterations have been observed to be elevated in apparently healthy workers exposed to benzene3-6, indicating the critical roles of cytogentic changes in benzene-induced leukemogenesis. 
Generally, a single FISH assay examines only one or a few whole chromosomes or specific loci per slide, so multiple hybridizations need to be conducted on multiple slides to cover all of the human chromosomes. Spectral karyotyping (SKY) allows visualization of the whole genome simultaneously, but the requirement for special software and equipment limits its application7. Here, we describe a novel FISH assay, OctoChrome-FISH, which can be applied for Chromosomics, which we define here as the simultaneous analysis of all 24 human chromosomes on one slide in human studies, such as chromosome-wide aneuploidy study (CWAS)8. The basis of the method, marketed by Cytocell as the Chromoprobe Multiprobe System, is an OctoChrome device that is divided into 8 squares, each of which carries three different whole chromosome painting probes (Figure 1). Each of the three probes is directly labeled with a different colored fluorophore, green (FITC), red (Texas Red), and blue (Coumarin). The arrangement of chromosome combinations on the OctoChrome device has been designed to facilitate the identification of the non-random structural chromosome alterations (translocations) found in the most common leukemias and lymphomas, for instance t(9;22), t(15;17), t(8;21), t(14;18)9. Moreover, numerical changes (aneuploidy) in chromosomes can be detected concurrently. The corresponding template slide is also divided into 8 squares onto which metaphase spreads are bound (Figure 2), and is positioned over the OctoChrome device. The probes and target DNA are denatured at high-temperature and hybridized in a humid chamber, and then all 24 human chromosomes can be visualized simultaneously. 
OctoChrome FISH is a promising technique for the clinical diagnosis of leukemia and lymphoma and for detection of aneuploidies in all chromosomes. We have applied this new Chromosomic approach in a CWAS study of benzene-exposed Chinese workers8,10.

A novel fluorescence in situ hybridization (FISH) method that simultaneously examines both numerical and structural chromosome alterations, particularly the specific chromosomal translocations associated with leukemia and lymphoma, of all 24 human chromosomes on a single device in one hybridization, is described.

在所有人类染色体24 chromosomics：检测的数值模拟和结构上的改动同时使用一种新型OctoChrome鱼含量

Fluorescence in situ hybridization (FISH) is a technique that allows specific DNA sequences to be detected on metaphase or interphase chromosomes in cell ...

Assessment of GFP Expression and Viability Using the Tali Image-Based Cytometer

Single-cell and population information are commonly obtained either by flow cytometry or fluorescence microscopy. However, these two methods provide different information. Flow cytometry gives quantitative multi-parametric information about physical characteristics and staining or expression, but doesn't allow for visualization. Stand-alone fluorescence microscopy provides visual data, but doesn't allow for straightforward quantitative measurements1.
Image-based cytometry bridges the gap between these two methods, enabling the quick visualization and simultaneous quantitative analysis of thousands of cells in heterogeneous populations2. Here, we present a method for performing cell viability and green fluorescent protein (GFP) expression assays using the Tali Image-Based Cytometer3. The Tali instrument is a 3-channel (bright field, green fluorescence, red fluorescence) benchtop assay platform that offers several advantages over flow cytometry and fluorescence microscopy. The Tali cytometer is less expensive, takes up less bench space, requires less maintenance, and the work flow has been simplified so that the operation and analysis is much simpler and quicker. The Tali cytometer is capable of performing a range of suspension cell-based assays, including GFP and red fluorescent protein (RFP) expression, apoptosis4-6 and cell viability analysis with propidium iodide (PI)7-11.
Here, we demonstrate the use of the Tali instrument in performing a cell viability assay in cells expressing GFP. GFP-transduced cells are stained using the Tali Viability Kit - Dead Cell Red. The cells are then pipetted into a Tali Cellular Analysis Slide and loaded into the cytometer. Bright field, red fluorescence and green fluorescence images are captured and analyzed using assay specific algorithms. Histograms are then generated to display cell size, PI fluorescence intensity, and GFP fluorescence intensity. These parameters can then be thresholded to home in on a specific cell population.
A side-by side comparison of the Tali Image-Based Cytometer and traditional flow cytometry demonstrates that the two methods provide comparable data regarding cell viability and protein expression. However, the Tali instrument provides additional visual information about the cell population that cannot be obtained using a flow cytometer.

This protocol describes how to perform cell viability and fluorescence expression assays using the Tali Image-Based Cytometer.

GFP的表达和使用的大里基于图像流式细胞仪的可行性评估

Single-cell and population information are commonly obtained either by flow cytometry or fluorescence microscopy. However, these two methods provide ...