Desarrollo de hardware fue apoyado por los contratos NNX09CA44C y NNX10CA97C NASA SBIR. Análisis de los datos de las manifestaciones de bloques y cargador de muestras ópticas fue apoyado por la NASA Fase III NNC11CA04C Contrato. La colección de la sangre humana se realizó utilizando NASA IRB Protocolo # SA-10-008. Software de control / adquisición proporcionada a través del Programa de Becas de Dispositivos Médicos de National Instruments. Moldes para los microchips se realizaron en las instalaciones de microfabricación Johns Hopkins y el Centro Harvard para sistemas a nanoescala. Otto J. Briner y Lucas Jaffe (Instituto de Medicina del ADN) ayudaron en el montaje en rack durante el verano de 2010. El personal de vuelo de la NASA de vídeo proporcionado imágenes de vídeo durante la semana de vuelo. Carlos Barrientos (Instituto de Medicina del ADN) proporcionó fotografía y cifra la asistencia. Un agradecimiento especial a la facilitó el acceso a la tecnología espacial para el Medio Ambiente 2010 del programa, la Oficina de la NASA Gravedad Reducida, la División de Contramedidas Adaptación Humana y del Centro de Investigación Glenn de la NASA,ZIN Technologies, y el Programa de Investigación en Seres Humanos.

<ol>
	<li>Thomas, R. A., Krishan, A., Robinson, D. M., Sams, C., Costa, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NASA/American+Cancer+Society+High-Resolution+Flow+Cytometry+Project-I.">NASA/American Cancer Society High-Resolution Flow Cytometry Project-I.</a> Cytometry. 43, 2-11 (2001).</li><li>Wen, J., Krishan, A., Thomas, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NASA/American+Cancer+Society+High-Resolution+Flow+Cytometry+Project+-+II.+Effect+of+pH+and+DAPI+concentration+on+dual+parametric+analysis+of+DNA/DAPI+fluorescence+and+electronic+nuclear+volume.">NASA/American Cancer Society High-Resolution Flow Cytometry Project - II. Effect of pH and DAPI concentration on dual parametric analysis of DNA/DAPI fluorescence and electronic nuclear volume.</a> Cytometry. 43, 12-15 (2001).</li><li>Krishan, A., Wen, J., Thomas, R. A., Sridhar, K. S., Smith, W. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NASA/American+Cancer+Society+High-Resolution+Flow+Cytometry+Project+-+III.+Multiparametric+analysis+of+DNA+content+and+electronic+nuclear+volume+in+human+solid+tumors.">NASA/American Cancer Society High-Resolution Flow Cytometry Project - III. Multiparametric analysis of DNA content and electronic nuclear volume in human solid tumors.</a> Cytometry. 43, 16-22 (2001).</li><li>Cram, L. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spin-offs+from+the+NASA+space+program+for+tumor+diagnosis.">Spin-offs from the NASA space program for tumor diagnosis.</a> Cytometry. 43, 1 (2001).</li><li>Crucian, B., Sams, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduced+gravity+evaluation+of+potential+spaceflight-compatible+flow+cytometer+technology.">Reduced gravity evaluation of potential spaceflight-compatible flow cytometer technology.</a> Cytometry B Clin. Cytom. 66 (1), 1-9 (2005).</li><li>Shi, W., Kasdan, H. L., Fridge, A., Tai, Y. -. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Four-part+differential+leukocyte+count+using+&#956;flow+cytometer.">Four-part differential leukocyte count using &#956;flow cytometer.</a> 2010 IEEE 23rd International Conference on Micro Electro Mechanical Systems. 13 (7), 1019-1022 (2010).</li><li>Tai, Y. -. C., Ho, C. -. M., Kasdan, H. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> In-Flight Blood Analysis Technology for Astronaut Health Monitoring NASA Human Research Program Investigators&#8217; Workshop. , (2010).</li><li>Shi, W., Guo, L. W., Kasdan, H., Fridge, A., Tai, Y. -. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Leukocyte+5-part+differential+count+using+a+microfluidic+cytometer.">Leukocyte 5-part differential count using a microfluidic cytometer.</a> 2011 16th International Solid-State Sensors, Actuators and Microsystems Conference. , 2956-2959 (2011).</li><li>Shi, W., Guo, L., Kasdan, H., Tai, Y. -. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Four-part+leukocyte+differential+count+based+on+sheathless+microflow+cytometer+and+fluorescent+dye+assay.">Four-part leukocyte differential count based on sheathless microflow cytometer and fluorescent dye assay.</a> Lab Chip. 13 (7), 1257-1265 (2013).</li><li>Dubeau-Laram&#233;e, G., Rivi&#232;re, C., Jean, I., Mermut, O., Cohen, L. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microflow1,+a+sheathless+fiber-optic+flow+cytometry+biomedical+platform:+Demonstration+onboard+the+international+space+station.">Microflow1, a sheathless fiber-optic flow cytometry biomedical platform: Demonstration onboard the international space station.</a> Cytometry A. , (2013).</li><li>Crucian, B., Quiriarte, H., Guess, T., Ploutz-Snyder, R., McMonigal, K., Sams, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Miniaturized+Analyzer+Capable+of+White-Blood-Cell+and+Differential+Analyses+During+Spaceflight.">A Miniaturized Analyzer Capable of White-Blood-Cell and Differential Analyses During Spaceflight.</a> Lab Medicine. 44 (4), 304-331 (2013).</li><li>Rehnberg, L., Russomano, T., Falc&#227;o, F., Campos, F., Everts, S. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+a+novel+basic+life+support+method+in+simulated+microgravity.">Evaluation of a novel basic life support method in simulated microgravity.</a> Aviat. Space. Environ. Med. 82 (2), 104-110 (2011).</li><li>Pump, B., Videbaek, R., Gabrielsen, A., Norsk, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterial+pressure+in+humans+during+weightlessness+induced+by+parabolic+flights.">Arterial pressure in humans during weightlessness induced by parabolic flights.</a> J. Appl. Physiol. 87 (3), 928-932 (1999).</li><li>Strauch, S. M., Richter, P., Schuster, M., H&#228;der, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+beating+pattern+of+the+flagellum+of+Euglena+gracilis+under+altered+gravity+during+parabolic+flights.">The beating pattern of the flagellum of Euglena gracilis under altered gravity during parabolic flights.</a> J. Plant Physiol. 167 (1), 41-46 (2010).</li><li>Sams, C. F., Crucian, B. E., Clift, V. L., Meinelt, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+whole+blood+staining+device+for+use+during+space+shuttle+flights.">Development of a whole blood staining device for use during space shuttle flights.</a> Cytometry. 37 (1), 74-80 (1999).</li><li>Smith, S. M., Davis-Street, J. E., Fontenot, T. B., Lane, 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=Assessment+of+a+portable+clinical+blood+analyzer+during+space+flight.">Assessment of a portable clinical blood analyzer during space flight.</a> Clin. Chem. 43, 1056-1065 (1997).</li><li>Weigl, B. H., Kriebel, J., Mayes, K. J., Bui, T., Yager, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole+Blood+Diagnostics+in+Standard+Gravity+and+Microgravity+by+Use+of+Microfluidic+Structures+(T-Sensors).">Whole Blood Diagnostics in Standard Gravity and Microgravity by Use of Microfluidic Structures (T-Sensors).</a> Microchimica Acta. 131 (1-2), 75-83 (1999).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revolutionizing+Medical+Technology+for+Earth+and+Space.">Revolutionizing Medical Technology for Earth and Space.</a> Canadian Space Agency. , (2012).</li><li>Peytavi, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+device+for+rapid+(&lt;15+min)+automated+microarray+hybridization.">Microfluidic device for rapid (&lt;15 min) automated microarray hybridization.</a> Clin. Chem. 51, 1836-1844 (2005).</li><li>Groemer, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+feasibility+of+laryngoscope-guided+tracheal+intubation+in+microgravity+during+parabolic+flight:+a+comparison+of+two+techniques.">The feasibility of laryngoscope-guided tracheal intubation in microgravity during parabolic flight: a comparison of two techniques.</a> Anesthesia and analgesia. 101 (5), 1533-1535 (2005).</li><li>Johnston, S. L., Campbell, M. R., Billica, R. D., Gilmore, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiopulmonary+resuscitation+in+microgravity:+efficacy+in+the+swine+during+parabolic+flight.">Cardiopulmonary resuscitation in microgravity: efficacy in the swine during parabolic flight.</a> Aviat. Space Environ. Med. 75 (6), 546-550 (2004).</li><li>Panait, L., Broderick, T., Rafiq, A., Speich, J., Doarn, C. R., Merrell, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+laparoscopic+skills+in+microgravity+anticipates+the+space+surgeon.">Measurement of laparoscopic skills in microgravity anticipates the space surgeon.</a> Am. J. Surg. 188 (5), 549-552 (2004).</li><li>Kirkpatrick, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraperitoneal+gas+insufflation+will+be+required+for+laparoscopic+visualization+in+space:+a+comparison+of+laparoscopic+techniques+in+weightlessness.">Intraperitoneal gas insufflation will be required for laparoscopic visualization in space: a comparison of laparoscopic techniques in weightlessness.</a> J. Am. Coll. Surg. 209 (2), 233-241 (2009).</li><li>Campbell, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endoscopic+surgery+in+weightlessness:+the+investigation+of+basic+principles+for+surgery+in+space.">Endoscopic surgery in weightlessness: the investigation of basic principles for surgery in space.</a> Surg. Endosc. 15 (12), 1413-1418 (2001).</li><li>Caiani, E. G., Sugeng, L., Weinert, L., Capderou, A., Lang, R. M., Va&#239;da, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Objective+evaluation+of+changes+in+left+ventricular+and+atrial+volumes+during+parabolic+flight+using+real-time+three-dimensional+echocardiography.">Objective evaluation of changes in left ventricular and atrial volumes during parabolic flight using real-time three-dimensional echocardiography.</a> J. Appl. Physiol. 101 (2), 460-468 (2006).</li><li>Ansari, R., Manuel, F. K., Geiser, M., Moret, F., Messer, R. K., King, J. F., Suh, K. I., Manns, F., S derberg, P. G., Ho, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+choroidal+blood+flow+in+zero+gravity.">Measurement of choroidal blood flow in zero gravity.</a> Ophthalmic technologies XII : 19-20 January 2002, San Jose, USA. , 177-184 (2002).</li><li>Foldager, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+venous+pressure+in+humans+during+microgravity.">Central venous pressure in humans during microgravity.</a> J. Appl. Physiol. 81 (1), 408-412 (1996).</li><li>Hausmann, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytosolic+calcium,+hydrogen+peroxide+and+related+gene+expression+and+protein+modulation+in+Arabidopsis+thaliana+cell+cultures+respond+immediately+to+altered+gravitation:+parabolic+flight+data.">Cytosolic calcium, hydrogen peroxide and related gene expression and protein modulation in Arabidopsis thaliana cell cultures respond immediately to altered gravitation: parabolic flight data.</a> Plant Biol. (Stuttg). 16 (1), 120-128 (2014).</li><li>Thiel, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+alterations+of+cell+cycle+control+proteins+in+human+T+lymphocytes+in+microgravity).">Rapid alterations of cell cycle control proteins in human T lymphocytes in microgravity).</a> Cell Commun. Signal. 10 (1), 1 (2012).</li><li>Tsuda, T., Kitagawa, S., Yamamoto, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimation+of+electrophoretic+mobilities+of+red+blood+cells+in+1-G+and+microgravity+using+a+miniature+capillary+electrophoresis+unit.">Estimation of electrophoretic mobilities of red blood cells in 1-G and microgravity using a miniature capillary electrophoresis unit.</a> Electrophoresis. 23, 2035-2039 (2002).</li><li>Paul, A. -. L., Manak, M. S., Mayfield, J. D., Reyes, M. F., Gurley, W. B., Ferl, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parabolic+flight+induces+changes+in+gene+expression+patterns+in+Arabidopsis+thaliana.">Parabolic flight induces changes in gene expression patterns in Arabidopsis thaliana.</a> Astrobiology. 11 (8), 743-758 (2011).</li><li>Zeredo, J. L., Toda, K., Matsuura, M., Kumei, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+responses+to+partial-gravity+conditions+in+rats.">Behavioral responses to partial-gravity conditions in rats.</a> Neurosci. Lett. 529 (2), 108-111 (2012).</li><li>Taube, J. S., Stackman, R. W., Calton, J. L., Oman, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+head+direction+cell+responses+in+zero-gravity+parabolic+flight.">Rat head direction cell responses in zero-gravity parabolic flight.</a> J. Neurophysiol. 92 (5), 2887-2897 (2004).</li><li>Hilbig, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+altered+gravity+on+the+swimming+behaviour+of+fish.">Effects of altered gravity on the swimming behaviour of fish.</a> Adv. Space Res. 30 (4), 835-841 (2002).</li><li>Yang, J., Qi, L., Chen, Y., Ma, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+Fabrication+of+a+Three+Dimensional+Spiral+Micromixer.">Design and Fabrication of a Three Dimensional Spiral Micromixer.</a> Chinese J. Chem. 31, 209-214 (2013).</li><li>Zhang, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Realization+of+planar+mixing+by+chaotic+velocity+in+microfluidics.">Realization of planar mixing by chaotic velocity in microfluidics.</a> Microelectron. Eng. 88, 959-963 (2011).</li><li>Liu, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Passive+mixing+in+a+three-dimensional+serpentine+microchannel.">Passive mixing in a three-dimensional serpentine microchannel.</a> J. Microelectromechanical Syst. 9, 190-197 (2000).</li></ol>

Eugene Y. Chan, Candice Bae, y Julia Z. Sharpe son inventores de patentes de tecnología relacionados presentadas a través del Instituto de Medicina de ADN, una entidad comercial.

El método descrito aquí permitió demostración efectiva de los principales componentes de la tecnología (la carga de la muestra, de mezcla de microfluidos, y detección óptica) durante los 2010 RÁPIDOS vuelos parabólicos, con resultados comparables a pruebas en tierra. Los métodos de formación y SOP descritos aquí fueron particularmente eficaz, y ayudaron a iluminar herramientas y otro ser &#39;muletas&#39; confiado en para demostraciones prácticas que no estarían disponibles a bordo del vuelo parabólico. <...

La insuficiencia de los diagnósticos de salud en el espacio listo actuales presenta un factor limitante para profundizar la exploración espacial tripulada. Diagnóstico necesitan ser integral, fácil de usar en gravedad reducida y relativamente poco afectada por las tensiones de lanzamiento y el vuelo espacial (por ejemplo, altas fuerzas G, la vibración, la radiación, los cambios de temperatura y presión de la cabina cambios). Los avances en las pruebas de punto de atención (POCT) pueden traducirse en soluciones efectivas de vuelo espacial a través del uso de las muestras de pacientes más pequeños (por ejemplo, un pinchazo en el dedo), fluidos más simples y de menor tamaño (es decir, de microfluidos), y la reducción de los requerimientos de energía eléctrica, entre otros ventajas. La citometría de flujo es un enfoque atractivo para POC en el espacio debido a la amplia utilidad de la tecnología, incluyendo hacia el recuento de células y cuantificación de biomarcadores, así como la miniaturización potencial significativo. Anterior citómetros de flujo en el espacio relevante incluyen la &#39;efic embalaje nucleariency &#39;(NPE) instrumento que utiliza la fluorescencia de arco de la lámpara simultánea inducida y volumen electrónico (Coulter volumen) de medición 1-4, un relativamente pequeño flujo de sobremesa citómetro de representación de la&#39; primera generación de flujo en tiempo real de datos de citometría durante gravedad cero &#39;5, un &#39;microflujo sin vaina citómetro&#39; capaz de 4 y 5 parte de glóbulos blancos (WBC) recuento diferencial utilizando pretratado 5 l muestras de sangre entera de 6-9, y un &#39;fibra óptica basada en&#39; citómetro de flujo recientemente probado a bordo en el Internacional Estación Espacial 10. La evaluación de la tecnología de diagnóstico para aplicaciones espaciales potenciales se realiza normalmente a bordo de aeronaves en gravedad reducida que utilizan una trayectoria de vuelo aproximadamente parabólica para simular un nivel deseado de ingravidez (por ejemplo, gravedad cero, marciana-gravedad) 11. La evaluación es un reto porque las oportunidades de vuelo son limitados, repetventanas cortas sitivos de la microgravedad puede hacer que sea difícil evaluar metodologías o procesos que normalmente requieren períodos ininterrumpidos más largos de 20 a 40 segundos, y manifestaciones pueden requerir equipo adicional no utilizado fácilmente en vuelo 12-15. Por otra parte, las manifestaciones anteriores de tecnologías de diagnóstico in vitro (IVD) usados ​​en, o diseñados para, gravedad reducida son limitadas y mucho trabajo permanece inédito. Además de los citómetros de flujo anteriormente, otros IVD-tecnologías espaciales relevantes descritos en la literatura incluyen un dispositivo de tinción de sangre entera para aplicaciones de inmunofenotipaje 16, un sistema automatizado de cámara basado en citómetro de 12, un analizador clínico de mano para potenciometría integrado, amperometría, y Conductometría 12,17, un dispositivo de microfluidos &#39;T-sensor&#39; para la cuantificación del analito que se basa en una mezcla basada en la difusión y la separación 18, y una rotación &#39;laboratorio en un CD&#39; plataforma de diagnóstico 19,20. Los recién llegados a las pruebas de gravedad reducida también pueden mirar a demostraciones de vuelo parabólico no relacionadas con el diagnóstico in vitro cuando se trata de hacer de la evaluación dispositivo posible (o averiguar lo que es posible). Las manifestaciones de otros experimentos médicos o biológicos anterior con la preparación del vuelo, las estrategias durante el vuelo, y el equipo de pruebas de vuelo bien documentado se incluyen en la Tabla 1 15, 21-35. Estos pueden ser de carácter informativo debido a la inclusión de tareas manuales en vuelo, el uso de equipo especializado, y la contención experimental. <table border="0" cellpadding="0" cellspacing="0"><tbody><tr height="17"><td height="17" style="height:17px;width:133px;"> Categoría </td><td style="width:497px;"> Ejemplos </td></tr><tr height="40"><td height="40" style="height:40px;width:133px;"> Atención médica de emergencia </td><td style="width:497px;"> La intubación traqueal (laringoscopio guiada, en maniken) 21, de soporte vital cardíaco (cerdos anestesiados) 22 </td></tr><tr height="20"><td height="20" style="height:20px;width:133px;"> La atención quirúrgica </td><td style="width:497px;"> La cirugía laparoscópica (vídeo simula 23, en cerdos anestesiados 24,25) </td></tr><tr height="41"><td height="41" style="height:41px;width:133px;"> Imagen médica o evaluación fisiología </td><td style="width:497px;"> Ultrasonido con cámara de presión negativa inferior del cuerpo 26, medidor de flujo Doppler (montado cabeza) 27, monitor de la presión venosa central 28 </td></tr><tr height="65"><td height="65" style="height:65px;width:133px;"> Equipos biológica especializada </td><td style="width:497px;"> Lector de microplacas (y la caja en vuelo guante) 29, sistema de control de temperatura para los experimentos del ciclo celular 30, microscopio (campo claro, contraste de fase y fluorescencia multicanal capaz) 15, capilarunidad de electroforesis acoplado al microscopio de vídeo 31 </td></tr><tr height="20"><td height="20" style="height:20px;width:133px;"> Otro </td><td style="width:497px;"> Recolección de plantas con fórceps 32, contenida ratas 33,34 y 35 de peces para la observación </td></tr></tbody></table> Tabla 1. Antena vuelo de demostración con ejemplos bien descrito Métodos / Experimentos Para ampliar los ejemplos anteriores y proporcionar un mayor conocimiento de exitosas demostraciones en vuelo, estamos presentando un procedimiento modular y adaptable para la construcción y operación de un prototipo citómetro de flujo con la tecnología de mezcla de microfluidos relacionada como parte de un banco de pruebas de vuelo parabólico. La plataforma permite a las manifestaciones de la carga de la muestra, la mezcla de microfluidos, y la detección de las partículas fluorescentes, y se puso a prueba a bordo de la NASA de 2010 facilitó el acceso a Medio Ambiente Espacial (FAST) flig parabólicaHTS, volados del 29 de septiembre al 1 de octubre de 2010. Estas demostraciones tirar desde el principio, medio y final, respectivamente, de un potencial de flujo de trabajo de dispositivo en el que se cargan, diluidos o mezclados con reactivos de muestras de sangre por punción capilar-tamaño, y se analizaron por medio de óptica detección. Escalar un citómetro de flujo en una unidad compacta requiere innovación y la selección cuidadosa parte. Personalizado y componentes fuera de la plataforma se usan aquí, elegido como mejores aproximaciones iniciales de elecciones finales de componentes, y pueden ser adaptables a los designios de otros innovadores. Siguiendo un esquema de opciones de componentes prototipo, la configuración se describe en una estructura de soporte que sirve de esqueleto para el montaje de perforación. Componentes del prototipo se asignan lugares, asegurados, y acompañados por los componentes adicionales necesarios para la experimentación con éxito. La atención se desplaza a procedimientos más abstractas que implican un procedimiento estándar de operación (SOP) el desarrollo, la capacitación y otros aspectos logísticos. Por último, los procedimientos de demostración-específica sondescrito. Las estrategias que aquí se describen y las opciones de apoyo a los componentes del equipo de perforación (por ejemplo, un microscopio, caja de acrílico, etc.), aunque implementado aquí por prototipo específico, hablan a las cuestiones generales y desafíos relevantes para probar cualquier equipo de diagnóstico en sangre en un ambiente de gravedad reducida . En los vuelos de 2010, dos lunar-gravedad (el logro de aproximadamente 1/6 gravedad de la tierra) y dos vuelos de microgravedad fueron programadas a través de 4 días, aunque en última instancia, estos fueron reprogramados en 3 días. Las manifestaciones se llevaron a cabo a bordo de un avión de pasajeros del jet modificado de gestión privada, de fuselaje estrecho 36. Cada vuelo, a condición 30-40 parábolas, cada uno produciendo aproximadamente 20 seg de alta gravedad (aproximadamente 1,8 g), seguido de 20-25 seg de las condiciones de gravedad reducida. Después se ejecuta la mitad de las parábolas, el avión se detuvo por un período de alrededor de 5-10 minutos en el nivel de vuelo para que el avión para dar la vuelta y regresar hacia el lugar de aterrizaje, mientras que performing el resto de las parábolas.

<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

Hasta hace poco, se recogieron muestras de sangre de los astronautas durante el vuelo, transportados a la Tierra en el transbordador espacial, y se analizaron en los laboratorios terrestres. Si los seres humanos son a viajar más allá de la órbita baja de la Tierra, una transición hacia el espacio listo, punto-de-cuidado se requiere (POC) de pruebas. Este tipo de pruebas tiene que ser integral, fácil de realizar en un ambiente de gravedad reducida, y no afectada por las tensiones de lanzamiento y el vuelo espacial. Dispositivos POC Innumerables han sido desarrollados para imitar homólogos a escala de laboratorio, pero la mayoría tienen aplicaciones estrechas y pocos tienen uso demostrable en un ambiente reducido la gravedad en vuelo. De hecho, las manifestaciones de los diagnósticos biomédicos en gravedad reducida se limitan por completo, por lo que la elección de los componentes y ciertos desafíos logísticos difícil acercarse al tratar de probar la nueva tecnología. Para ayudar a llenar el vacío, presentamos un método modular para la construcción y operación de un dispositivo de diagnóstico de sangre prototipo y su p asociadoarabolic banco de pruebas de vuelo que cumple con las normas para las pruebas de vuelo a bordo de un vuelo parabólico, aviones reducido la gravedad. El primer método se centra en el montaje de equipo de perforación, pruebas en vuelo reducido la gravedad de un citómetro de flujo y un chip de microfluidos mezcla compañero. Los componentes son adaptables a otros diseños y algunos componentes personalizados, tales como un cargador de muestras microvolumen y la micromezclador pueden ser de particular interés. El método entonces enfoque cambia a la preparación del vuelo, ofreciendo pautas y sugerencias para prepararse para una prueba de vuelo con éxito en lo que respecta a la formación de los usuarios, el desarrollo de un procedimiento operativo estándar (SOP), y otras cuestiones. Por último, se describen en vuelo los procedimientos experimentales específicas a nuestras manifestaciones.

Los resultados representativos para la demostración micromezclador aparecen en la Figura 7, como se ve por la cámara CCD montada en el microscopio estereoscópico. Mezcla puede evaluarse visualmente en cualquier punto a lo largo de la espiral, así como en el canal de salida para los experimentos que involucran dos conjuntos de fluidos: sangre / solución salina y de colorante azul / amarillo. El análisis cuantitativo de las imágenes de dos dimensiones puede incluir la determinación de la uniformid...

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

Diagnósticos sanguíneos vuelo espacial necesitan innovación. Pocas manifestaciones han sido publicados ilustrando en vuelo, la tecnología de diagnóstico de salud de gravedad reducida. Aquí presentamos un método para la construcción y operación de un banco de pruebas de vuelo parabólico para un diseño de la citometría de flujo en el punto de atención de prototipo, con componentes y estrategias de preparación adaptables a otras configuraciones.

Reducción de gravedad Demostraciones entorno de hardware de un prototipo miniaturizado citómetro de flujo y Compañero Microfluidos Tecnología de mezcla

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 vistas.  DNA Medicine Institute.  Diagnósticos sanguíneos vuelo espacial necesitan innovación. Pocas manifestaciones han sido publicados ilustrando en vuelo, la tecnología de diagnóstico de salud de gravedad reducida. Aquí presentamos un método para la construcción y operación de un banco de pruebas de vuelo parabólico para un diseño de la citometría de flujo en el punto de atención de prototipo, con componentes y estrategias de preparación adaptables a otras configuraciones. 

Video: 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.

Un dispositivo de microfluidos con los patrones de Surco para el estudio del comportamiento celular

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

Investigación

Biología

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.

Un dispositivo de microfluidos gradiente de generación de Biología Celular

 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-linfocitos capturar usando un chip de microfluidos desechables para el VIH

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.

Un compartimiento de Multi-CNS neuronas-glia Co-cultivo de microfluidos Plataforma

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.

La sonda de microfluidos: Operación y Uso de tratamiento de superficies localizadas

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.

Adhesión y agregación plaquetaria bajo flujo con células de microfluidos de flujo

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.

Microfluidos Co-cultivo de células epiteliales y bacterias para la Investigación de la señal mediada Soluble Interacciones

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.

Cuantificación de mezcla utilizando imágenes por resonancia magnética

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.

Chromosomics: Detección de alteraciones numéricas y estructurales en todos los 24 cromosomas humanos de manera simultánea utilizando un nuevo ensayo FISH OctoChrome

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.

Evaluación de la expresión de GFP y viabilidad mediante el Tali basada en imágenes citómetro

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