Sviluppo hardware è stato sostenuto dalla NASA SBIR Contratti NNX09CA44C e NNX10CA97C. L&#39;analisi dei dati per il blocco e il campione del caricatore dimostrazioni ottiche è stato sostenuto dalla NASA Fase III Contratto NNC11CA04C. La raccolta del sangue umano è stata effettuata utilizzando la NASA IRB protocollo # SA-10-008. Software di controllo / acquisizione fornito attraverso il National Instruments Medical Device Grant Program. Stampi per i microchip sono stati effettuati presso l&#39;impianto di microfabbricazione Johns Hopkins e il Centro di Harvard per Nanoscale Systems. Otto J. Briner e Luca Jaffe (DNA Istituto di Medicina) aiutati nel montaggio del rack durante l&#39;estate del 2010. La NASA personale di volo di video riprese video fornito durante la settimana di volo. Carlos Barrientos (DNA Medicine Institute) ha fornito assistenza fotografia e la figura. Un ringraziamento speciale al agevolato accesso allo spazio dell&#39;ambiente per Technology 2010 Programma, la gravità Ufficio NASA ridotto, l&#39;adeguamento umana e contromisure Division, NASA Glenn Research Center,ZIN Technologies, e il programma di ricerca umano.

<ol>
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Eugene Y. Chan, Candice Bae, e Julia Z. Sharpe sono gli inventori di brevetti tecnologici relativi depositati attraverso l&#39;Istituto di Medicina del DNA, un ente commerciale.

Il metodo qui descritto ha permesso efficace dimostrazione delle principali componenti della tecnologia (campione di carico, miscelazione microfluidica, e di rivelazione ottica) nel corso del 2010 voli parabolici veloce, con risultati paragonabili a prove a terra. Formazione e SOP metodi qui descritti sono stati particolarmente efficaci, e hanno contribuito a illuminare strumenti e delle altre &#39;stampelle&#39; invocata per le dimostrazioni pratiche che non sarebbero disponibili a bordo del volo parabolico. <p cla...

L&#39;inadeguatezza della diagnostica attuali di salute spazio-ready presenta un fattore limitante per una più profonda esplorazione spaziale con equipaggio. Diagnostica devono essere completo, facile da usare in gravità ridotta, e relativamente inalterata alle sollecitazioni di lancio e il volo spaziale (ad esempio, g-forze elevate, vibrazioni, radiazioni, sbalzi di temperatura, e pressione cabina modifiche). Gli sviluppi nei test point-of-care (POCT) possono tradursi in soluzioni efficaci volo spaziale attraverso l&#39;uso di campioni dei pazienti più piccoli (ad esempio, una puntura dito), fluidica più semplici e più piccole (ad esempio, microfluidica), e ridotto fabbisogno di energia elettrica, tra l&#39;altro vantaggi. Citometria a flusso è un approccio interessante per in-spazio POC causa della ampia utilità della tecnologia, compresa verso conteggio delle cellule e biomarker quantificazione, nonché un significativo potenziale miniaturizzazione. Spazio rilevante Precedente flusso citometri includono il &#39;effic imballaggio nucleareiency &#39;(NPE), strumento utilizzato fluorescenza simultanea ad arco lampada indotta e volume elettronico (Coulter volume) di misura 1-4, un relativamente piccolo flusso da banco citometro che rappresenta la&#39; prima generazione di flusso in tempo reale dei dati durante la citometria a gravità zero &#39;5, un &#39;microflusso guaina citometro&#39; in grado di globuli bianchi (WBC) conta differenziale a 4 e 5 parti utilizzando pretrattato 5 ml campioni di sangue intero 6-9, e &#39;in fibra ottica a base di&#39; citofluorimetro recentemente testato a bordo in International Stazione Spaziale 10. Valutare tecnologia diagnostica per le applicazioni spaziali potenziali viene in genere eseguita aerei ridotta gravità che utilizzano una traiettoria di volo di circa parabolica per simulare un livello prescelto di assenza di peso (ad esempio, a gravità zero, marziano-gravità) 11 a bordo. La valutazione è impegnativo, perché opportunità di volo sono limitati, repetbrevi finestre itive di microgravità può rendere difficile la valutazione delle metodologie o processi che normalmente richiedono periodi ininterrotti più lunghi di 20-40 sec, e le dimostrazioni possono richiedere ulteriori attrezzature non facilmente utilizzato in volo 12-15. Inoltre, precedenti dimostrazioni di tecnologie (IVD) in vitro diagnostici usati in, o progettati per, gravità ridotta sono limitati e molto lavoro resta inedito. Oltre ai citofluorimetri sopra, altri IVD-tecnologie spaziali rilevanti descritti in letteratura comprendono un dispositivo complesso colorazione sangue per applicazioni immunofenotipizzazione 16, un sistema automatico fotocamera basata citometro 12, un analizzatore clinico palmare per potenziometria integrato, amperometria e conductometry 12,17, un dispositivo di microfluidica &#39;T-sensor&#39; per la quantificazione degli analiti che si basa sulla miscelazione basato sulla diffusione e la separazione 18, e la rotazione di un &#39;laboratorio su un CD&#39; piattaforma diagnostica 19,20. I nuovi arrivati ​​alla sperimentazione gravità ridotta possono anche guardare a dimostrazioni di volo parabolico estranei alla diagnostica in vitro quando si tenta di rendere possibile la valutazione del dispositivo (o capire ciò che è possibile). Dimostrazioni di altri precedenti esperimenti medici o biologici con la preparazione del volo, le strategie in volo, e le apparecchiature di prova di volo ben documentata sono incluse nella tabella 1 15, 21-35. Questi possono essere informativo a causa dell&#39;inclusione di manuale in volo compiti, l&#39;uso di attrezzature specializzate e di contenimento sperimentale. <table border="0" cellpadding="0" cellspacing="0"><tbody><tr height="17"><td height="17" style="height:17px;width:133px;"> Categoria </td><td style="width:497px;"> Esempi </td></tr><tr height="40"><td height="40" style="height:40px;width:133px;"> Assistenza medica di emergenza </td><td style="width:497px;"> L&#39;intubazione tracheale (laringoscopio-guidato, su Manika) 21, supporto vitale cardiaco (suini anestetizzati) 22 </td></tr><tr height="20"><td height="20" style="height:20px;width:133px;"> Cure chirurgiche </td><td style="width:497px;"> La chirurgia laparoscopica (video simulato 23, in maiali anestetizzati 24,25) </td></tr><tr height="41"><td height="41" style="height:41px;width:133px;"> Medical imaging o la valutazione fisiologia </td><td style="width:497px;"> Ultrasuoni con camera a pressione negativa inferiore del corpo 26, Doppler flussometro (montato la testa) 27, di controllo della pressione venosa centrale 28 </td></tr><tr height="65"><td height="65" style="height:65px;width:133px;"> Attrezzature biologico Specialized </td><td style="width:497px;"> Lettore di micropiastre (e vano portaoggetti in volo) 29, sistema di controllo della temperatura per gli esperimenti del ciclo cellulare 30, microscopio (campo chiaro, contrasto di fase, e multi-canale in grado di fluorescenza) 15, capillareUnità di elettroforesi accoppiato al video microscopio 31 </td></tr><tr height="20"><td height="20" style="height:20px;width:133px;"> Altro </td><td style="width:497px;"> Raccolta delle piante con una pinza 32, contenuta ratti 33,34 e pesce 35 per l&#39;osservazione </td></tr></tbody></table> Tabella 1. Antenna volo dimostrativo Esempi con ben descritto Metodi / Esperimenti Per espandere su esempi precedenti e di fornire una maggiore comprensione di successo dimostrazioni in volo, vi presentiamo una procedura modulare e adattabile per la costruzione e il funzionamento di un prototipo citofluorimetro con relativa tecnologia di miscelazione microfluidica come parte di un test di volo parabolico rig. L&#39;impianto di perforazione permette dimostrazioni di caricamento del campione, la miscelazione microfluidica, e rivelazione di particelle fluorescenti, ed è stato testato a bordo 2010 NASA agevolato l&#39;accesso allo spazio ambiente (FAST) flig parabolicahts, volato dal 29 settembre al 1 ° ottobre 2010. Queste dimostrazioni tirare dal inizio, metà e fine, rispettivamente, di un flusso di lavoro dispositivo di potenziale in cui campioni di sangue dal polpastrello dimensioni sono caricati, diluiti o miscelati con i reagenti, e analizzati con ottica rilevamento. Scalare un citofluorimetro in un&#39;unità compatta richiede innovazione e un&#39;attenta selezione parte. Personalizzato e componenti off-the-shelf sono usati qui, scelto come migliori approssimazioni iniziali di scelte componente finale, e possono essere adattabili ai disegni di altri innovatori. In seguito una descrizione dei componenti scelte prototipo configurazione è descritto su una struttura di supporto che funge da scheletro per assemblaggio rig. Componenti prototipi vengono assegnati punti, fissati e accompagnati da componenti aggiuntivi necessari per la sperimentazione di successo. L&#39;attenzione si sposta quindi a procedure più astratte che coinvolgono procedure operative standard (SOP) lo sviluppo, la formazione, e altri aspetti logistici. Infine, le procedure specifiche per dimostrazione sonodescritto. Le strategie qui descritte e le scelte di elementi portanti di rig (ad esempio, il microscopio, scatola acrilica, ecc), anche se attuato qui per prototipo specifico, parlano le questioni generali e le sfide rilevanti per la verifica qualsiasi apparecchiatura diagnostica di sangue in un ambiente a gravità ridotta . Nel 2010 Voli, due lunare-gravità (il raggiungimento di circa 1/6 della gravità terrestre) e due voli di micro-gravità sono stati programmati in 4 giorni, anche se in ultima analisi, questi sono stati riprogrammati in 3 giorni. Le dimostrazioni sono state effettuate a bordo di una modifica gestite da privati, narrow-body jet aereo di linea 36. Ogni volo, a condizione 30-40 parabole, ciascuna producendo circa 20 sec di alta gravità (circa 1,8 g), seguita da 20-25 secondi di condizioni di ridotta gravità. Dopo mezzo delle parabole furono eseguiti, il piano in pausa per un periodo di circa 5-10 minuti in volo livellato per consentire il piano di girarsi e tornare verso il luogo di destinazione, mentre performing il resto delle parabole.

<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

Fino a poco tempo, i campioni di sangue sono stati raccolti astronauti in volo, trasportati a terra sullo Space Shuttle, e analizzati nei laboratori terrestri. Se gli esseri umani devono viaggiare oltre l&#39;orbita terrestre bassa, una transizione verso lo spazio-ready, point-of-care è richiesto (POC) test. Tale test deve essere completo, facile da eseguire in un ambiente a gravità ridotta, e non influenzata dalle sollecitazioni di lancio e il volo spaziale. Innumerevoli dispositivi POC sono stati sviluppati per imitare controparti scala di laboratorio, ma la maggior parte hanno le applicazioni strette e pochi hanno l&#39;uso dimostrabile in un ambiente a gravità ridotta in volo. In realtà, dimostrazioni di diagnostica biomedica in gravità ridotta sono limitati del tutto, rendendo la scelta dei componenti e di alcuni problemi logistici difficile avvicinarsi quando si cerca di testare nuove tecnologie. Per contribuire a colmare il vuoto, vi presentiamo un metodo modulare per la costruzione e il funzionamento di un dispositivo diagnostico prototipo di sangue e il suo p associatoarabolic banco di prova di volo che soddisfano gli standard per prove di volo a bordo di un volo parabolico, aerei ridotta gravità. Il metodo si concentra sulla prima assemblea rig per in-volo, test ridotta gravità di un citofluorimetro e un compagno microfluidica chip di miscelazione. I componenti sono adattabili ad altri disegni e alcuni componenti personalizzati, come ad esempio una pala campione microvolumi e micromixer possono essere di particolare interesse. Il metodo quindi attenzione si sposta alla preparazione del volo, offrendo linee guida e suggerimenti per la preparazione per un test di volo di successo per quanto riguarda la formazione degli utenti, lo sviluppo di una procedura operativa standard (SOP), e altre questioni. Infine, sono descritti in volo procedure sperimentali specifiche per le nostre manifestazioni.

Risultati rappresentativi per la dimostrazione micromixer appaiono in figura 7, come visto dalla telecamera CCD montato stereomicroscopio. Miscelazione può essere valutata visivamente in qualsiasi punto lungo la spirale, così come nel canale di uscita per esperimenti di due serie di fluidi: sangue / salini e blu / colorante giallo. Analisi quantitativa delle immagini bidimensionali può includere la determinazione di uniformità ombra su tutta la larghezza del canale in diverse regioni, come mostrato ...

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

Diagnostica del sangue di volo spaziale hanno bisogno di innovazione. Poche manifestazioni sono stati pubblicati illustrando in volo, tecnologia diagnostica sanitaria ridotta gravità. Qui vi presentiamo un metodo per la costruzione e l&#39;esercizio di un impianto parabolica test di volo di un prototipo di design point-of-care citometria a flusso, con componenti e strategie di preparazione adattabili ad altre impostazioni.

Ridotto gravità ambiente hardware Dimostrazioni di un prototipo miniaturizzato Citofluorimetro e Companion Microfluidic tecnologia di miscelazione

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 Views.  DNA Medicine Institute.  Diagnostica del sangue di volo spaziale hanno bisogno di innovazione. Poche manifestazioni sono stati pubblicati illustrando in volo, tecnologia diagnostica sanitaria ridotta gravità. Qui vi presentiamo un metodo per la costruzione e l'esercizio di un impianto parabolica test di volo di un prototipo di design point-of-care citometria a flusso, con componenti e strategie di preparazione adattabili ad altre impostazioni. 

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 a microfluidi con pattern Groove per studiare il comportamento cellulare

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

Ricerca

Biologia

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 gradiente di generazione del dispositivo Microfluidic di Biologia Cellulare

 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-linfociti Capture utilizzando un chip monouso Microfluidic per l&#39;HIV

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 multi-vano SNC Neuron-glia Co-cultura Microfluidic piattaforma

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 Microfluidic: funzionamento e utilizzo per l&#39;elaborazione delle superfici localizzate

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.

L&#39;adesione e aggregazione piastrinica in condizioni di flusso utilizzando celle di flusso Microfluidic

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.

Microfluidica Co-coltura di cellule epiteliali e batteri per le indagini solubile del segnale mediato da Interazioni

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.

Quantificare miscelazione con risonanza magnetica

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: Rilevamento delle alterazioni numeriche e strutturali in tutti i 24 cromosomi umani simultaneamente utilizzando un nuovo metodo di analisi 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.

Valutazione di espressione GFP e sostenibilità Utilizzando la Image-Based Tali Citometro

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