Bu çalışma Bilim ve Teknoloji Tayvan Bakanlığı&#39;ndan hibe tarafından kısmen desteklenmiştir (EN 104-2628-e-007-001-My3 (CMC)), ve Taichung Gaziler General Hospital <span style="color: rgb(34, 34, 34); font-family: arial, sans-serif; font-size: 12.8px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: 1; word-spacing: 0px; -webkit-text-stroke-width: 0px; display: inline !important; float: none; background-color: rgb(255, 255, 255);">(TCVGH-1056904C (MYH)).

DİKKAT: Uygun laboratuvar hijyen uygulamaları gerekmektedir. Bu POC cihazı kullanırken eldiven ve evrensel önlemler gereklidir. Yeterli sterilizasyon prosedürleri düzgün yapılmadığı takdirde sonuçları veya enfeksiyon bulaşma meydana gelebilir. 1. Test Şerit Cihazları hazırlayın <ol><li> Temas açısı ölçümü 8 (Şekil 3) ile temizlik pamuk dış tabakasının hidrofobikliğini belirleyin. </li><li> Bir kağıt kesici (Şekil 2A) ile 5.5 cm x 1 cm parçalar halinde temizlik pamuk keserek pamuk bazlı analitik cihaz imal. </li><li> Üç satır laminasyon filmi standart bir levha delik açın (ø = 0.5 cm); birbirini takip eden iki delik arasındaki mesafe 1 cm&#39;dir. NOT: Her laminasyon filmi, standart bir tabaka (5.5 cm x 2 cm) (Şekil 2B) kullanılarak yapılabilir üç test alanları gibi bir çok 36 nolu ayrı ayrı test şeritleri. </li><li> temizlik pamuk Sandwich (characterizLaminasyon filmi, örneğin, bir tarafta laminasyon filmi delik delinmiş sac, ve diğer tarafta da laminasyon filmi bir delinmemiş tabaka (deliksiz) iki adet arasında dış hidrofobiklik ve iç hidrofilikliği) tarafından ifa. Cihazı paketlemek için bir laminar içine derleme yerleştirin. Bu cihazın mekanik mukavemet artırır ve harici, su geçirmez katmanı sağlar. NOT: Delik çapları 0.47 cm laminasyon sonra 0.5 cm ~ den küçülecek. Bazı hafif boşluklar her deliğin çevresinde temizlik pamuk ve laminasyon filmi arasında kalacaktır. Bu hafif bir boşluk, bir sonraki aşamada yapılan deney tampon çevrelerinde yerleştirilmesini kolaylaştırır. </li><li> Yani montaj, bireysel 5,5 cm x 1 cm test şeridi cihazları kesmek için makas bir çift kullanın, delinmiş laminasyon sac / pamuk ped / deliksiz laminasyon sac &quot;sandviç&quot; (Şekil 2C). (Daha sonra açıklanacağı şekilde 48 cihazlar standart eğrileri oluşturmak için toplam 18 cihazlar yapmakNitrit deneyi için, BSA Deney 15 aygıtları ve ürobilinojen tahlilinde 15 cihazları). </li><li> Her test alanının ortasına (laminasyon filmi kapsamına girmeyen) den (küçük bir akışkan kanalı gibi) bir yarık kesmek için keskin bir bıçak ya da jilet kullanın, karşısında ve boyuna cihazın hidrofilik tabakasına test alanının hidrofobik katmanı sayesinde, dikkatli olmak sandviç cihazı (Şekil 2B) derinliği ile yarım kesmek için. </li><li> Laminasyon (Şekil 2E) her deliğin çevresine boşlukları içine kaydırarak kağıt testi yastıkları (ø = 0.5 cm) uygulayın. </li></ol> 2. idrar tahlili için Standartlar ve Gösterge Çözümleri hazırlayın <ol><li> (Di) deiyonize su, 100 ml 69.0 mg sodyum nitrit çözülmesiyle 10.0 mm nitrit stok çözelti hazırlayın. Bir nitrit konsantrasyonu, standart solüsyonların dizi (2.500, 1.250, 625, 312, 156, ve 78 mcM) oluşturmak için DI su nitrit stok solüsyonu sulandırmak (Şekil4A). </li><li> 100 ml PBS çözeltisi 415 mg BSA çözülmesiyle 60 uM sığır serum albümini (BSA) bir stok çözelti hazırlayın (10 M fosfat tamponu, 0.0027 M potasyum klorür ve 0.137 M sodyum klorür, pH 7.0). BSA konsantrasyonu standart solüsyonların bir dizi oluşturmak için DI su BSA stok solüsyonu sulandırmak (30, 15, 7.5, 3.75 ve 1.875 uM) (Şekil 4B). </li><li> 100 ml Dİ su içinde 2.5 gr urobilinogen çözülmesiyle 25 g / L ürobilinojen stok çözelti hazırlayın. Ürobilinojen konsantrasyonu, standart çözümlerin (7.8, 15.6, 31.2, 62.5 ve 125 mg / L) (Şekil 4C) bir dizi oluşturmak için DI su stok ürobilinojen çözeltisi ile seyreltilir. </li><li> 50 mM sülfanilamid, 330 mM sitrik asit ve 10 mM N- (1-naftil) etilenediamin diklorid içeren nitrit gösterge solüsyonu hazırlanır. </li><li> 250 mM sitrat tampon çözeltisi (pH 1.8) ve% 95 etanol içinde tetrabromophenol mavi 3.3 mM solüsyonu BSA gösterge solüsyonu hazırlanır. </li><li>0.1 M 4-dimetilamin benzaldehit içeren ürobilinojen indikatör çözeltisi hazırlayın. </li></ol> 3. Gösterge Pads hazırlayın <ol><li> Üç yuvarlak şekilli pedleri veya diskler (ø = 0.5 cm) kromatografi kağıttan kesilmiş ve bir çözüm standart bir eğri (bölüm 4) oluşturmak için her test şeridi cihaz üzerine onlara (üç deney şeridi üzerinde her bir deney için bir tane) her yerleştirin. </li><li> toplanma üç laminasyon filmi deliklerin her anda, laminasyon filmi ve kesim deliğin çok kenarlarında pamuk test pedi arasında hafif bir boşluk görüyoruz. laminasyon filmi delik kağıt test pedi daha çapı biraz daha küçüktür, dikkat edin. NOT: Birlikte, bu özellikler kağıt testi ped cihazının pamuk testi ped kısmına karşı uygulanacak izin böylece laminasyon filmi deliğin kenarlarına altında bahsedilen ince boşluk, yani içine kağıt testi ped kayma kenarları. Bu yerde kağıt testi yastıkları tutar. </li><li> Nitrit assa hazırlayınCihaz y Porsiyon. <ol><li> Coat nitrit göstergesi (adım 2.4 hazırlanmış) çözeltisi ve ortam sıcaklığında tamamen kurumasını bekleyin 4 ul nitrit tahlil ped. </li></ol></li><li> Tespit Cihazı Toplam Protein Deney Porsiyon hazırlayın. <ol><li> Kat BSA göstergesi çözeltisi (adım 2.5 hazırlandı) ve oda sıcaklığında tam olarak kurumasına izin 4 ul toplam protein tahlili pedi. </li></ol></li><li> Tespit Cihazının Uroblinogen Deneyi Porsiyon hazırlayın. <ol><li> Kat uroblinogen (adım 2.6 hazırlandı) ve oda sıcaklığında tam olarak kurumasına izin 4 ul uroblinogen deney takımı. </li></ol></li></ol> 4. Pamuk temelli Analitik Cihaz için standart eğrileri oluşturun <ol><li> hazırlanan her standart için, önceden hazırlanmış bir cihazın emici ucunu batırın ~ 10 sn (emilim hacmi standart çözeltisi içine (deney göstergelerinin (bakınız bölüm 3.4) ile önceden yüklenmiş)200-300 ul) (Şekil 2F). </li><li> 10 dk kolorimetrik reaksiyonlar (Şekil 2F) tamamlamak için bir çevre ortamında pamuk tabanlı bir aygıt yerleştirin. üç kopya halinde her bir deney tekrar edin. </li><li> Her test pedi (Şekil 2G) meydana gelen kolorimetrik değişim analiz cihazı tarayın. Bir masaüstü tarayıcı ve görüntü çözünürlüğü 600 dpi RGB modunda tarama testi pedleri camına deney yastıkları ortaya çıkan bir yer. jpeg formatında her görüntü kaydetme. </li><li> Ilgili renk yoğunlukları (Şekil 2H) belirlemek için görüntü analizi yazılımı kullanılarak bir bilgisayarda taranan görüntüyü analiz edin. <ol><li> http://imagej.nih.gov/ij/ at ImageJ yazılımı indirin. Taranan görüntü dosyaları açın. Menü komutları Görüntü seçin ve 8-bit gri tonlama RGB görüntüleri dönüştürmek için &quot;Type&quot; ı seçin. </li><li> Test pedi analitik alanı seçmek için araç çubuğundaki &quot;Oval&quot; aracını kullanın. Seçin menü comm içinde &quot;Analiz&quot;Eleştiri ve renk yoğunluğunu analiz etmek &quot;Tedbir&quot; seçeneğini seçin. Kayıt yoğunluğu sonuçlarını anlamına gelir. </li></ol></li><li> Nitrit, total protein ve ürobilinojen deneyleri (Şekil 2I) standart eğrileri kurmak. <ol><li> Standart nitrit, toplam protein ve ürobilinojen çözeltiler farklı konsantrasyonlarını gösterir ve stastical yazılımı kullanılarak, iki boyutlu bir koordinat sistemine göre ortalama yoğunluğu sonuçlarıyla karşılaştırıldı. </li><li> karesel model ve lineer regresyon modelleri ile total protein ve ürobilinojen deneyleri için standart eğrileri tarafından nitrit tayini için standart eğriye uyacak. </li></ol></li></ol> 5. Kuruluş Standart Eğri değerleri kullanma Bilinmeyen Numune Konsantrasyonları belirleyin <ol><li> , Nitrit, total protein ve bilinmeyen çözümler ürobilinojen konsantrasyonlarını belirlemek standart çözüm konsantrasyonlarını ve nitrit yerine ölçülen yoğunluk sonuçları, total protein ölçmek için tarif edildiği gibi cihazı kullanmak ve urobi için2. adımda inşa modellerin denklem içine linogen deneyleri. </li></ol>

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The authors declare that they have no competing financial interests. S.C.L., C.M.K., F.G.T., and C.M.C. are inventors of a patent (patent No.: US 8691162 B1) owned by National Tsing Hua University on the basis of the work reported in this manuscript. S.C.L., C.M.K., F.G.T., and C.M.C.have patents pending in Taiwan and China based on the work reported in this manuscript. M.Y.H. declares no potential conflict of interest.

Bu protokol kritik adım ve filtre kağıdı (kromatografisi filtre kağıdı veya kantitatif filtre kağıdı) (hidrofobiklik / hidrofiliklik değişen) pamuklu malzeme uygun kombinasyonunu belirlemek dahildir. İyi planlanmış ve idam cihaz tasarımı en iyi performansı kolorimetrik deneyleri için niteliklerini vermektedir. Bizim kolorimetrik tahlil sonuçlarından, burada sunulan pamuk bazlı analitik cihaz birden hastalık tespiti için bir platform olarak büyük bir potansiyele gösteriyor. <p class="jove_c...

Nokta-bakım gelişimi (POC), uygun fiyatlı, sağlam ve kolay kullanılan küresel sağlık 1,2 geliştirmek için zorunludur olan tanı cihazları. Özellikle, cihazlar selüloz yüzeylerde (örneğin, kağıt, iplik ve pamuk) nedeniyle aynı anda her yerde, uygun fiyatta, hızlı sonuçlar 3-7 sağlamak için kullanım, sağlamlık ve kapasite kolaylığı, düşük maliyet analizi için umut verici analitik platformlar sağlamak oluşan. Burada, idrar tahlili için bir yanal akış tabanlı biçimini kullanan bir pamuk bazlı analitik cihazın geliştirilmesini açıklayacak. Bu pamuk bazlı analitik cihaz birkaç anahtar avantajları ile alternatif bir algılama yaklaşım sağlamaktadır: en az insan çaba ile i) üretimi; ii) düşük maliyetli; iii) kapasitesi çapraz kontaminasyon endişeleri olmaksızın birden çok farklı deneyleri yapmak için kullanılacak; . iv) aygıt bağımsızlığı, yani, yetenek ek donanım ve / veya elektrik olmadan çalıştırılmak üzere; ve h) Hızı (kolorimetrik tahlilleri) 10 dakika içinde tamamlanabilir. Bu pamuk bazlı analitik cihazı yapısı dört bölüme ayrılabilir: i) dış hidrofobik katman üzerinde doğal olarak hidrofobik olan pamuk; ii) dahili olarak hidrofiliktir ve sıvı fitilleme bir ulaşım kanalı olarak hizmet veren pamuk, bağlanan ve pamuk kullanılan ancak reaksiyon / test pedleri yerleştirilmesi için delikleri delinmiş içeren sıkıştırır iii) laminasyon filmi; ve iv) kolorimetrik deney için reaksiyon alanları olarak pamuk (spesifik olarak, boşlukta laminasyon filmi üzerinden delinmiş) dış yüzeyi üzerine yerleştirilen / kaplanmış Reaktif ile gömülürler kromatografi kağıdı test pedleri, (örneğin, nitrit, toplam protein, pH ve ürobilinojen tahlilleri) ve sonuçları ekran. aşağıdaki şekilde test mekanizmadır. pamuk bazlı analitik cihaz borç yoluyla yarım nüfuz hatları ile atılırsaPamuk malzemesinin h numunesi reaktif pedler kullanılan ulaşmasını sağlayan bir akış kanalı oluşturmak için. Çözelti testi pedleri (Şekil 1) emme ucundan akışkan kanalı boyunca kötü olduğunu bunun üzerine analitik cihazın emici kenarı, hedef numunenin içine batırılır. Test yastığının emici gücü pamuktan daha büyük olduğundan, deney yastıkları tarafından emilir çözümler hiçbir reflow akışkan kanala geri kalmayacak şekilde sıkıca Test ped kağıt içinde yer alır ve kolorimetrik sonuçlar daha sonra sabit hale Test pedi malzemesi. Reaksiyon sonunda, kolorimetrik sonuçlar bir masaüstü tarayıcısı üzerinden kaydedilir ve görüntü analiz yazılımı ile analiz edilmiştir.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>bovine serum albumin</td>
			<td>Sigma-Aldrich, US</td>
			<td>No. 9048468</td>
			<td>&#8805; 99%</td>
		</tr>
		<tr>
			<td>nitrite&#160;</td>
			<td>Sigma-Aldrich, US</td>
			<td>No. 7632000</td>
			<td>&#8805; 99%</td>
		</tr>
		<tr>
			<td>urobilinogen&#160;</td>
			<td>Santa Cruz Bio, US</td>
			<td>No. SC-296690</td>
			<td></td>
		</tr>
		<tr>
			<td>citrate</td>
			<td>Sigma-Aldrich U.S</td>
			<td>No. 6132043</td>
			<td>&#8805; 99%</td>
		</tr>
		<tr>
			<td>tetrabromophenol blue</td>
			<td>Sigma-Aldrich U.S</td>
			<td>No. 4430255</td>
			<td>&#8805; 99%</td>
		</tr>
		<tr>
			<td>sulfanilamide</td>
			<td>Sigma-Aldrich U.S</td>
			<td>No. 63741</td>
			<td>&#8805; 99%</td>
		</tr>
		<tr>
			<td>citric acid&#160;</td>
			<td>Sigma-Aldrich U.S</td>
			<td>No. 77929</td>
			<td>&#8805; 99.5%</td>
		</tr>
		<tr>
			<td>&#160;N-(1-naphthyl) ethylenediamine dihydrochloride</td>
			<td>Sigma-Aldrich U.S</td>
			<td>No. 1465254</td>
			<td>&#8805; 98%</td>
		</tr>
		<tr>
			<td>4-(Dimethylamine)benzaldehyde</td>
			<td>AlfaAesar, U.S</td>
			<td>No. A11712</td>
			<td>&#8805; 98%</td>
		</tr>
		<tr>
			<td>Methyl Red sodium salt</td>
			<td>sigma, U.S</td>
			<td>No. 114502</td>
			<td>&#8805;95%</td>
		</tr>
		<tr>
			<td>Bromothyle blue</td>
			<td>sigma, U.S</td>
			<td>No. 114413</td>
			<td>&#8805;95%</td>
		</tr>
		<tr>
			<td>Shiseido Cleansing Cotton</td>
			<td>Shiseido, Japan</td>
			<td>No. 79014</td>
			<td></td>
		</tr>
		<tr>
			<td>chromatography paper</td>
			<td>GE Healthcare Whatman, Springfield Mill, UK</td>
			<td>No. 30306132</td>
			<td></td>
		</tr>
		<tr>
			<td>plastic substrate</td>
			<td>lamination film, MAS</td>
			<td>A4</td>
			<td>216 mm &#215; 303 mm</td>
		</tr>
		<tr>
			<td>scanner</td>
			<td>microtek scanmaker</td>
			<td>&#160;i2400</td>
			<td></td>
		</tr>
		<tr>
			<td>paper cutter</td>
			<td>Life paper cutter</td>
			<td>No.306</td>
			<td></td>
		</tr>
		<tr>
			<td>laminator</td>
			<td>AURORA&#160;</td>
			<td>LM4231H</td>
			<td></td>
		</tr>
		<tr>
			<td>laminator film</td>
			<td>UNI LAMI&#160;</td>
			<td>4A</td>
			<td></td>
		</tr>
	</tbody>
</table>

fabricating cotton analytical devices

Sağlam, düşük maliyetli analitik cihaz kullanıcı dostu, hızlı ve uygun fiyatlı olmalıdır. Bu tür cihazlar da kıt örnekleri ile çalışır ve takip tedavi için bilgi sağlamak gerekir. Burada, bir yanal akış tabanlı bir format kullanır ve kolay, hızlı, ucuz imal edilir ve yürütmek için kullanılabilir (yani, nitrit, toplam protein ve ürobilinojen tahlilleri) değerlendirme cihazı pamuk bazlı idrar tahlili gelişimini göstermektedir çapraz bulaşma endişeleri olmaksızın çoklu testler. Pamuk akımı bazlı analiz için kaldıraçlı doğal emme özellikleri olan selüloz liflerinin oluşmaktadır. Bizim pamuk bazlı analitik cihazın sade ama şık fabrikasyon süreci bu çalışmada açıklanmıştır. Pamuk yapısı ve test pedi düzenlenmesi hidrofobikliği ve her malzemenin emme gücü yararlanır. Çünkü bu fiziksel özelliklerinin, kolorimetrik sonuçları ısrarla testin yapışabilirped. Bu cihaz zamanında klinik bilgi almak için hekimlerin sağlar ve erken müdahale için bir araç olarak büyük bir potansiyel gösterir.

Biz başarıyla hidrofil (iç kısım) ile karakterize piyasada mevcut temizlik pamuk ve hidrofobik (dış kısım) özellikleri (Şekil 1A) kullanarak pamuk bazlı analitik cihazların geliştirilmesini göstermiştir. 3 temas açısı ölçüm sonuçlarını göstermektedir. Dış pamuk hidrofobik arayüzü 127,35 ° ± 4.73 ° idi. (Şekil 2F) analizleri kullanıcı dostu bir bakış açısıyla, burada kullanılan ko...

Watch this Scientific Journal Video about Fabricating Cotton Analytical Devices at JoVE.com

Fabricating Cotton Analytical Devices

To investigate simple fabrication approaches for multiple assay needs, we created a fluid-absorbing channel system made of cotton material. This device was used to establish a multiple detection platform, and solve contamination issues that commonly affect lateral flow-based biomedical devices, for clinical urinalysis of nitrite, total protein, and urobilinogen.

Pamuk Analitik Cihazlar imalatı

A robust, low-cost analytical device should be user-friendly, rapid, and affordable. Such devices should also be able to operate with scarce samples and ...

Research

JoVE Journal

Bioengineering

6.6K Görüntüleme.  National Tsing Hua University. To investigate simple fabrication approaches for multiple assay needs, we created a fluid-absorbing channel system made of cotton material. This device was used to establish a multiple detection platform, and solve contamination issues that commonly affect lateral flow-based biomedical devices, for clinical urinalysis of nitrite, total protein, and urobilinogen.

Video: Fabricating Cotton Analytical Devices

Manufacturing Devices and Instruments for Easier Rat Liver Transplantation

Orthotopic rat liver transplantation is a popular model, which has been shown in a recent JoVE paper with the use of the "quick-linker" device. This technique allows for easier venous cuff-anatomoses after a reasonable learning curve. The device is composed of two handles, which are carved out from scalpel blades, one approximator, which is obtained by modifying Kocher's forceps, and cuffs designed from fine-bore polyethylene tubing. The whole process can be performed at a low-cost using common laboratory material. The present report provides a step-by-step protocol for the design of the required pieces and includes stencils.

We describe the design of the &#x201C;quick-linker&#x201D; device for easier orthotopic rat liver transplantation.

Kolay Sıçan Karaciğer Nakli için üretim cihazlar ve Araçlar

Orthotopic rat liver transplantation is a popular model, which has been shown in a recent JoVE paper with the use of the "quick-linker" device. This ...

Biyomühendislik

Fabricating Complex Culture Substrates Using Robotic Microcontact Printing (R-&#181;CP) and Sequential Nucleophilic Substitution

In tissue engineering, it is desirable to exhibit spatial control of tissue morphology and cell fate in culture on the micron scale. Culture substrates presenting grafted poly(ethylene glycol) (PEG) brushes can be used to achieve this task by creating microscale, non-fouling and cell adhesion resistant regions as well as regions where cells participate in biospecific interactions with covalently tethered ligands. To engineer complex tissues using such substrates, it will be necessary to sequentially pattern multiple PEG brushes functionalized to confer differential bioactivities and aligned in microscale orientations that mimic in vivo niches. Microcontact printing (&#956;CP) is a versatile technique to pattern such grafted PEG brushes, but manual &#956;CP cannot be performed with microscale precision. Thus, we combined advanced robotics with soft-lithography techniques and emerging surface chemistry reactions to develop a robotic microcontact printing (R-&#956;CP)-assisted method for fabricating culture substrates with complex, microscale, and highly ordered patterns of PEG brushes presenting orthogonal &#8216;click&#8217; chemistries. Here, we describe in detail the workflow to manufacture such substrates.

Fabricating Complex Culture Substrates Using Robotic Microcontact Printing R- &#181;CP and Sequential Nucleophilic Substitution

Cell culture substrates functionalized with microscale patterns of biological ligands have immense utility in the field of tissue engineering. Here, we demonstrate the versatile and automated manufacture of tissue culture substrates with multiple, micropatterned poly(ethylene glycol) brushes presenting orthogonal chemistries that enable spatially precise and site-specific immobilization of biological ligands.

Robotik Microcontact Baskı (R-μCP) kullanarak Karmaşık Kültürü Substratlar ve Sıralı nükleofilik Giren Fabricating

In tissue engineering, it is desirable to exhibit spatial control of tissue morphology and cell fate in culture on the micron scale. Culture substrates ...

Generation of Scalable, Metallic High-Aspect Ratio Nanocomposites in a Biological Liquid Medium

The goal of this protocol is to describe the synthesis of two novel biocomposites with high-aspect ratio structures. The biocomposites consist of copper and cystine, with either copper nanoparticles (CNPs) or copper sulfate contributing the metallic component. Synthesis is carried out in liquid under biological conditions (37 &#176;C) and the self-assembled composites form after 24 hr. Once formed, these composites are highly stable in both liquid media and in a dried form. The composites scale from the nano- to micro- range in length, and from a few microns to 25 nm in diameter. Field emission scanning electron microscopy with energy dispersive X-ray spectroscopy (EDX) demonstrated that sulfur was present in the NP-derived linear structures, while it was absent from the starting CNP material, thus confirming cystine as the source of sulfur in the final nanocomposites. During synthesis of these linear nano- and micro-composites, a diverse range of lengths of structures is formed in the synthesis vessel. Sonication of the liquid mixture after synthesis was demonstrated to assist in controlling average size of the structures by diminishing the average length with increased time of sonication. Since the formed structures are highly stable, do not agglomerate, and are formed in liquid phase, centrifugation may also be used to assist in concentrating and segregating formed composites.

Here we present a protocol to synthesize novel, high-aspect ratio biocomposites under biological conditions and in liquid media. The biocomposites scale from nanometers to micrometers in diameter and length, respectively. Copper nanoparticles (CNPs) and copper sulfate combined with cystine are the key components for the synthesis.

Biyolojik Sıvı Ortamda Ölçeklenebilir, Metalik Yüksek Aspect Ratio Nanokompositlerin Üretimi

The goal of this protocol is to describe the synthesis of two novel biocomposites with high-aspect ratio structures. The biocomposites consist of copper ...

Fabricating Superhydrophobic Polymeric Materials for Biomedical Applications

Superhydrophobic materials, with surfaces possessing permanent or metastable non-wetted states, are of interest for a number of biomedical and industrial applications. Here we describe how electrospinning or electrospraying a polymer mixture containing a biodegradable, biocompatible aliphatic polyester (e.g., polycaprolactone and poly(lactide-co-glycolide)), as the major component, doped with a hydrophobic copolymer composed of the polyester and a stearate-modified poly(glycerol carbonate) affords a superhydrophobic biomaterial. The fabrication techniques of electrospinning or electrospraying provide the enhanced surface roughness and porosity on and within the fibers or the particles, respectively. The use of a low surface energy copolymer dopant that blends with the polyester and can be stably electrospun or electrosprayed affords these superhydrophobic materials. Important parameters such as fiber size, copolymer dopant composition and/or concentration, and their effects on wettability are discussed. This combination of polymer chemistry and process engineering affords a versatile approach to develop application-specific materials using scalable techniques, which are likely generalizable to a wider class of polymers for a variety of applications.

Two- and three-dimensional superhydrophobic polymeric materials are prepared by electrospinning or electrospraying biodegradable polymers blended with a lower surface energy polymer of similar composition.

Biyomedikal Uygulamalar için Süperhidrofobik Polimer Malzemeler imalatı

Superhydrophobic materials, with surfaces possessing permanent or metastable non-wetted states, are of interest for a number of biomedical and industrial ...

Using Adhesive Patterning to Construct 3D Paper Microfluidic Devices

We demonstrate the use of patterned aerosol adhesives to construct both planar and nonplanar 3D paper microfluidic devices. By spraying an aerosol adhesive through a metal stencil, the overall amount of adhesive used in assembling paper microfluidic devices can be significantly reduced. We show on a simple 4-layer planar paper microfluidic device that the optimal adhesive application technique and device construction style depends heavily on desired performance characteristics. By moderately increasing the overall area of a device, it is possible to dramatically decrease the wicking time and increase device success rates while also reducing the amount of adhesive required to keep the device together. Such adhesive application also causes the adhesive to form semi-permanent bonds instead of permanent bonds between paper layers, enabling single-use devices to be non-destructively disassembled after use. Nonplanar 3D origami devices also benefit from the semi-permanent bonds during folding, as it reduces the likelihood that unrelated faces may accidently stick together. Like planar devices, nonplanar structures see reduced wicking times with patterned adhesive application vs uniformly applied adhesive.

We demonstrate the use of patterned aerosol adhesives to construct 3D paper microfluidic devices. This method of adhesive application forms semi-permanent bonds between layers, enabling single-use devices to be non-destructively disassembled after use and to ease folding complex nonplanar structures.

3D Kağıt mikroakışkan Cihazlar Construct Yapışkan Desenlendirme kullanma

We demonstrate the use of patterned aerosol adhesives to construct both planar and nonplanar 3D paper microfluidic devices. By spraying an aerosol ...

High-throughput Identification of Bacteria Repellent Polymers for Medical Devices

Medical devices are often associated with hospital-acquired infections, which place enormous strain on patients and the healthcare system as well as contributing to antimicrobial resistance. One possible avenue for the reduction of device-associated infections is the identification of bacteria-repellent polymer coatings for these devices, which would prevent bacterial binding at the initial attachment step. A method for the identification of such repellent polymers, based on the parallel screening of hundreds of polymers using a microarray, is described here. This high-throughput method resulted in the identification of a range of promising polymers that resisted binding of various clinically relevant bacterial species individually and also as multi-species communities. One polymer, PA13 (poly(methylmethacrylate-co-dimethylacrylamide)), demonstrated significant reduction in attachment of a number of hospital isolates when coated onto two commercially available central venous catheters. The method described could be applied to identify polymers for a wide range of applications in which modification of bacterial attachment is important.

A high-throughput microarray method for the identification of polymers which reduce bacterial surface binding on medical devices is described.

Tıbbi Cihazlar Bakterilerin yüksek verimlilik Kimlik Savar Polimerler

Medical devices are often associated with hospital-acquired infections, which place enormous strain on patients and the healthcare system as well as ...

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Recent advances in modular DNA assembly techniques have enabled synthetic biologists to test significantly more of the available &#34;design space&#34; represented by &#34;devices&#34; created as combinations of individual genetic components. However, manual assembly of such large numbers of devices is time-intensive, error-prone, and costly. The increasing sophistication and scale of synthetic biology research necessitates an efficient, reproducible way to accommodate large-scale, complex, and high throughput device construction.
Here, a DNA assembly protocol using the Type-IIS restriction endonuclease based Modular Cloning (MoClo) technique is automated on two liquid-handling robotic platforms. Automated liquid-handling robots require careful, often times tedious optimization of pipetting parameters for liquids of different viscosities (e.g. enzymes, DNA, water, buffers), as well as explicit programming to ensure correct aspiration and dispensing of DNA parts and reagents. This makes manual script writing for complex assemblies just as problematic as manual DNA assembly, and necessitates a software tool that can automate script generation. To this end, we have developed a web-based software tool, http://mocloassembly.com, for generating combinatorial DNA device libraries from basic DNA parts uploaded as Genbank files. We provide access to the tool, and an export file from our liquid handler software which includes optimized liquid classes, labware parameters, and deck layout. All DNA parts used are available through Addgene, and their digital maps can be accessed via the Boston University BDC ICE Registry. Together, these elements provide a foundation for other organizations to automate modular cloning experiments and similar protocols.
The automated DNA assembly workflow presented here enables the repeatable, automated, high-throughput production of DNA devices, and reduces the risk of human error arising from repetitive manual pipetting. Sequencing data show the automated DNA assembly reactions generated from this workflow are ~95% correct and require as little as 4% as much hands-on time, compared to manual reaction preparation.

Here, an automated workflow to perform modular DNA &#34;device&#34; assembly using a modular cloning DNA assembly method on liquid-handling robots is presented. The protocol uses an intuitive software tool for generating liquid handler picklists for combinatorial DNA device library generation, which we demonstrate using two liquid handling platforms.

Robot sıvı işleme derleme modüler DNA cihazların otomatik

Recent advances in modular DNA assembly techniques have enabled synthetic biologists to test significantly more of the available &#34;design space&#34; ...

Multi-step Variable Height Photolithography for Valved Multilayer Microfluidic Devices

Microfluidic systems have enabled powerful new approaches to high-throughput biochemical and biological analysis. However, there remains a barrier to entry for non-specialists who would benefit greatly from the ability to develop their own microfluidic devices to address research questions. Particularly lacking has been the open dissemination of protocols related to photolithography, a key step in the development of a replica mold for the manufacture of polydimethylsiloxane (PDMS) devices. While the fabrication of single height silicon masters has been explored extensively in literature, fabrication steps for more complicated photolithography features necessary for many interesting device functionalities (such as feature rounding to make valve structures, multi-height single-mold patterning, or high aspect ratio definition) are often not explicitly outlined. 
Here, we provide a complete protocol for making multilayer microfluidic devices with valves and complex multi-height geometries, tunable for any application. These fabrication procedures are presented in the context of a microfluidic hydrogel bead synthesizer and demonstrate the production of droplets containing polyethylene glycol (PEG diacrylate) and a photoinitiator that can be polymerized into solid beads. This protocol and accompanying discussion provide a foundation of design principles and fabrication methods that enables development of a wide variety of microfluidic devices. The details included here should allow non-specialists to design and fabricate novel devices, thereby bringing a host of recently developed technologies to their most exciting applications in biological laboratories.

Multilayer microfluidic devices often involve the fabrication of master molds with complex geometries for functionality. This article presents a complete protocol for multi-step photolithography with valves and variable height features tunable to any application. As a demonstration, we fabricate a microfluidic droplet generator capable of producing hydrogel beads.

Ventilli Katmanlı mikroakışkan Cihazlar için çok adımlı Değişken Yükseklik Fotolitografi

Microfluidic systems have enabled powerful new approaches to high-throughput biochemical and biological analysis. However, there remains a barrier to ...

High-resolution Patterning Using Two Modes of Electrohydrodynamic Jet: Drop on Demand and Near-field Electrospinning

Electrohydrodynamic (EHD) jet printing has drawn attention in various fields because it can be used as a high-resolution and low-cost direct patterning tool. EHD printing uses a fluidic supplier to maintain the extruded meniscus by pushing the ink out of the nozzle tip. The electric field is then used to pull the meniscus down to the substrate to produce high-resolution patterns. Two modes of EHD printing have been used for fine patterning: continuous near-field electrospinning (NFES) and dot-based drop-on-demand (DOD) EHD printing. According to the printing modes, the requirements for the printing equipment and ink viscosity will differ. Even though two different modes can be implemented with a single EHD printer, the realization methods significantly differ in terms of ink, fluidic system, and driving voltage. Consequently, without a proper understanding of the jetting requirements and limitations, it is difficult to obtain the desired results. The purpose of this paper is to present a guideline so that inexperienced researchers can reduce the trial and error efforts to use the EHD jet for their specific research and development purposes. To demonstrate the fine-patterning implementation, we use Ag nanoparticle ink for the conductive patterning in the protocol. In addition, we also present the generalized printing guidelines that can be used for other types of ink for various fine-patterning applications.

Here, we present a protocol to produce high-resolution conductive patterns using electrohydrodynamic (EHD) jet printing. The protocol includes two modes of EHD jet printing: the continuous near-field electrospinning (NFES) and the dot-based drop-on-demand (DOD) EHD printing.

Yüksek çözünürlüklü Electrohydrodynamic Jet iki modu kullandıktan biçimlenme: damla talep ve yakın alan Electrospinning

Electrohydrodynamic (EHD) jet printing has drawn attention in various fields because it can be used as a high-resolution and low-cost direct patterning ...

Fabricating a Kidney Cortex Extracellular Matrix-Derived Hydrogel

Extracellular matrix (ECM) provides important biophysical and biochemical cues to maintain tissue homeostasis. Current synthetic hydrogels offer robust mechanical support for in vitro cell culture but lack the necessary protein and ligand composition to elicit physiological behavior from cells. This manuscript describes a fabrication method for a kidney cortex ECM-derived hydrogel with proper mechanical robustness and supportive biochemical composition. The hydrogel is fabricated by mechanically homogenizing and solubilizing decellularized human kidney cortex ECM. The matrix preserves native kidney cortex ECM protein ratios while also enabling gelation to physiological mechanical stiffnesses. The hydrogel serves as a substrate upon which kidney cortex-derived cells can be maintained under physiological conditions. Furthermore, the hydrogel composition can be manipulated to model a diseased environment which enables the future study of kidney diseases.

Here we present a protocol to fabricate a kidney cortex extracellular matrix-derived hydrogel to retain the native kidney extracellular matrix (ECM) structural and biochemical composition. The fabrication process and its applications are described. Finally, a perspective on using this hydrogel to support kidney-specific cellular and tissue regeneration and bioengineering is discussed.

Bir böbrek korteks ekstraselüler matrisi elde edilen hidrojel imalatı

Extracellular matrix (ECM) provides important biophysical and biochemical cues to maintain tissue homeostasis. Current synthetic hydrogels offer robust ...