This work was supported by the EC within the framework of the STIFF-FLOP FP7-ICT-2011.2.1 European Project (#287728).

Not: Bu protokol kanalı, tahrik boru hatları ve dış kılıf sertleşme, akışkan odaları içeren tek bir modül, fabrikasyonu aşamalarını anlatır. Aşağıdaki prosedür davlumbaz ve güvenlik nedenleriyle laboratuvar önlüğü ve eldiven giyme altında yürütülecek vardır. Daha önce belirtildiği gibi, elastomerik biriminin üretim süreci CAD yazılımı ile tasarlanmış kalıpların ardışık kullanım dayanmaktadır. Bunlar, Şekil 2&#39;de gösterildiği ve Tablo 1 &#39;de listelenen 13 parçadan oluşur. Silikon 1. Hazırlık <ol><li> A kısmının 12 g ve aynı plastik, cam veya Petri kabındaki B kısmının 12 g tartılır ve karıştırılarak, karıştırınız. Not: Malzeme oranlar iki bölümden oluşur, bu durumda, kullanılan özel silikon bağlı olarak değişebilir: Parça A (baz) ve kısım B (katalizörü). Bu oranı 1A kullanılır: 1B ağırlığı. </li><li> Karışımı içeren cam yerleştirin1 bar vakum basıncında bir gaz giderici makinesinde ed silikon malzemeler. Tüm kabarcıklar silikon malzemeden kaldırılır kadar vakum altında cam tutun. Kullanılan silikon için gaz alma işlemi yaklaşık 10 dakika sürer. Malzemeler kabarcıklarının varlığı tamamen arınmış olduğunda, makinenin içine atmosferik basınç ve geri yükleme silikon kullanabilirsiniz. </li></ol> Siliconic Modülü 2. Fabrikasyon <ol><li> Kalıp Meclisi. <ol><li> Sertleşme silindir ve cap_A (Şekil 3a) içine odaları üst yerleştirin. </li><li> Cap_A ikinci kat etrafında kabukları kapatın. </li></ol></li><li> İlk silikon döküm. <ol><li> Kapağın (Şekil 3b) kenarına monte edilmiş bir kalıbı up içinde silikon dökün. </li><li> Yaklaşık 30 dakika boyunca 60 ° C&#39;de bir fırın içinde kalıp yerleştirin. </li></ol></li><li> Kalıp yeniden düzenlenmesi. <ol><li> Dış kabukları çıkarın ve cap_A (Şekil 3c). </li><li> Odaları bazlar ve cap_B (Şekil 3d) içine sertleştirici silindirden silindir yerleştirin. </li><li> Yukarı doğru modülünün üst yüzeyi ile kapağın (Şekil 3e) kenarları arasında 10 mm&#39;lik bir boşluk olması için 10 mm kayar, modül üzerine yeniden kabukları kapatın. </li></ol></li><li> İkinci silikon döküm. <ol><li> Üst tarafı (yani, aynı zamanda sertleştirme silindir kadar) (Şekil 3f) kapsüllerinin kenarına yeniden düzenlenmiş kalıp up içinde silikon dökün. </li><li> Yaklaşık 30 dakika boyunca 60 ° C&#39;da bir fırın içine kalıp koyun. </li><li> Dış kabukları, cap_B ve (sertleştirme silindir hariç) odaları (Şekil 3g) sökün. </li></ol></li></ol> Tüpler 3. Yerleştirme <ol><li> Aynı istenilen uzunluğa (örneğin 300 mm) 3 tüpleri kesin. </li><li> Ob olmadan 10 mm, her borunun bir ucu etrafında siliconic tutkal koyuntüpler Structuring. </li><li> Siliconic birimi (Şekil 3h) kanalları adanmış 2 mm iç tüpler yerleştirin. </li><li> Oda sıcaklığında 12 dakikalık bir vulkanizasyon süresi izin verir ya da daha yüksek bir sıcaklıkta bir fırın içine modülü koymak - Kurutma işlemini hızlandırmak için (50 ° 60 °). </li></ol> Kıvrımlı Örgülü Kılıf 4. Fabrikasyon <ol><li> Genişletilebilir örgülü kılıf (modülün yaklaşık 15 kat yükseklik) 700 mm kesin. </li><li> Çapı 30 mm ve kabuğun içinde uzunluğu 250 mm metal silindir yerleştirin. </li><li> Aşağı doğru itin ve kıvrımın yaratmak amacıyla, silindir üzerinde kaydırarak kılıfı zorlar. </li><li> Bir kalıcı deformasyon elde edilene kadar mekanik 2-3 dakika boyunca 350 ° C&#39;de bir ısıtma tabancası ile bir kelepçe ve ısı ile yerine kılıf düzeltin. </li><li> Kılıf soğumasını ve iç silindir kaldırmak edelim. </li></ol> Dış Kılıf 5. Entegrasyon <ol><li> Geçcap_C deliklerinden borular. </li><li> Cap_C içine silikon 3 g dökün. </li><li> Çalışma düzlemi daha yüksek bir desteğe cap_C sıkıştırın. </li><li> Daha önce cap_C halinde imal modülün alt tarafı yerleştirin. </li><li> Modül etrafında kıvrılmış kılıfı kaydırın. </li><li> Cap_C içinde kılıfın ilk kıvrımın itin ve yeni dökülmüş silikon (Şekil 3i) içine batırın. </li><li> Yaklaşık 20 dakika boyunca 60 ° C&#39;da bir fırın içine kalıp koyun. </li><li> Cap_D (Şekil 3j) kullanılarak, üst tarafında kılıf düzeltmek için noktadan 5,1-5,6 aynı işlemi tekrarlayın. </li><li> Cap_C ve cap_D çıkarın. </li><li> Merkezi silindir (Şekil 3k) sökün. </li></ol> Granül Karıştırma Membran 6. Fabrikasyon <ol><li> Plastik bardak sıvı lateks 5 g dökün. </li><li> Yüzey kadar sıvı lateks içindeki membran (Şekil 2&#39;de gösterildiği gibi, son parça) için silindir daldırıntamamen kaplıdır. </li><li> 20 dakika boyunca bir başlık altında kurumaya bırakın. </li><li> Tekrarlayın noktalar 6.2 ve 6.3. </li><li> Kalıptan membran çıkarın. </li></ol> Granül Karıştırma Membran 7. Yerleştirme <ol><li> Bir boru arzu edilen uzunlukta (çapı 2 mm) (örneğin, 300 mm) kesin. </li><li> Yaklaşık 100 mm naylon 2 dokusunun bir kare parça kesin ve bir plastik film ya da parafın yapıştırıcı kullanarak bu doku ile tüpün bir ucunu kapatmak. </li><li> Kahve tozunun 4 g tartılır ve bir membran doldurun. </li><li> Dolu zarının içindeki tüp (filtreli sonu) yerleştirin ve bir plastik parafin film kullanarak tüp etrafında sabitleyin. </li><li> Tüp (membran sert olur) diğer tarafta bir vakum uygulayın. </li><li> Siliconic modülü (Şekil 3L) boş, merkezi kanal içinde membran yerleştirin. </li><li> Silikon modülüne sertleştirme zar uçlarını Tutkal. </li><li> Üst etrafında halkalar kapatınmodülü (Şekil 3m) yan. </li><li> Yüzey düzeltme amacıyla halkalar halinde silikonun 2 g dökün. </li><li> Kaputun altında ya da 60 ° &#39;de bir fırında silikon kurumasını bekleyin. </li><li> Halkalarını çıkartın. </li><li> Noktaları alt tarafı için 7.11 ile 7.8 (Şekil 3n) tekrarlayın. </li></ol>

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The authors have nothing to disclose.

The technique described in this protocol enables the fabrication of a pneumatically actuated soft unit usable for modular compliant structures. Thanks to the design of the molds and their simple assembly, it is possible to fabricate one complete module in about 4 hours with 7 main steps. The process of fabrication involves specific materials, which are easily available, and work should be carried out under a fume hood. An external set up including air valves, air compressor and vacuum pump is necessary to activate the mo...

Tıp alanında son trendler, cerrahi operasyonlar invazivlik bir azalma için bastırıyorlar. Minimal İnvazif Cerrahi (MIS) başarıyla karın operasyonlar için son birkaç yıl içinde geliştirilmiştir. MIS prosedürler karın duvarının üzerine yerleştirilen dört ya da beş erişim noktası (trokar) içinden araçların kullanımına dayanmaktadır. Trokar sayısını azaltmak amacıyla, araçlar Tek Bağlantı Laparoskopi (SPL) ya da Doğal Orifis endoskopik cerrahi (NOTES) 1 ile eklenebilir. Bu prosedürler, dış görünür izleri önlemek ancak cerrahi yürütülmesinde klinisyenler için zorluk artar. Bu sınırlamanın nedeni erişim azaltılmış puan ve önlemek veya organların yaklaşık 2 geçmek mümkün değildir aletleri, sert ve yarı sert doğa esas 3. Beceri ve hareketlilik kullanılarak geliştirilebilir eklemli ve hiper-gereksiz daha geniş ve daha karmaşık bir çalışma alanı kapsayabilir robotlar, incivücuttaki belirli bir hedef daha kolay 4, 5, 6, ulaşılacak ve 7 gerektiğinde geri çekme sistemleri olarak çalışmaya olanak. Esnek manipülatör böylece geleneksel araçlarla daha güvenli temas, doku uyumu artırabilir. Ancak, bu manipülatörler genellikle hedefe ulaşıldığında istikrar eksikliği ve genellikle çevredeki dokulara 8, temas kontrol edemez 9. Çalışmalar bu tür ahtapot kolu 10 ve fil gövde 11 olarak biyolojik yapılar üzerinde, son zamanlarda tasarımını ilham Bir Özgürlük (DOFs) ait Derecelerin gereksiz sayısı ve kontrol sertlik 12 ile esnek deforme olabilen ve uyumlu manipülatörler. Bu tip cihazların 13, 14, 15. Genel olarak, yumuşak ve esnek bir malzeme ile imal edilen, yüksek kuvvetler manipülatörler üretilmesini garanti etmezler pasif yaylar, akıllı materyaller, pnömatik elemanları veya tendonların kullanmaktadır. TO STIFF-FLOP (cerrahi işlemler için Rijitlik kontrol esnek ve Öğrenebilir manipülatör) manipülatör geçtiğimiz günlerde ahtapot yetenekleri esinlenerek NOTES ve SPL için yeni bir cerrahi aygıt olarak sunulmuştur. Önceki yumuşak manipülatörlerin sınırlamaları aşmak için, yüksek beceri, yüksek kuvvet ve kontrol sertlik 16 yanı sıra yumuşak bir gövdeye sahip. Manipülatör mimarisi modüler yaklaşıma dayanmaktadır: birden fazla üniteye, aynı yapı ve işlevleri ile birlikte entegre edilmiştir. Tek bir birim, Şekil 1 &#39;de gösterilmiştir. Bu çok fazlı bir imalat ile elde edilen bir elastomerik silindir üzerine dayanır. Kalıp ve döküm bileşenleri süreçlerinin montaj aşamaları (akışkan çalıştırılması için) üç boş odaları ve bir oyuk merkezi kanal 17 (konut için granül sıkışma tabanlı mekanizma 18) gömülü mümkün kılmaktadır. Odalar, böylece 120 ° &#39;de yer almaktadırir kombine enflasyon yönlü hareket ve uzama üretir. Buna ek olarak, harici bir örgülü kılıf böylece (eğilme ve uzama) modülü harekete odası harekete etkisi optimize, basınçlı zaman akışkan odaların dışa radyal genişlemesini sınırlamak için dışarıdan yerleştirilir. Orta kanal granüler malzeme ile doldurulmuş bir dış zar oluşan silindirik bir cihaz ev sahipliği yapmaktadır. Bir vakum basıncı uygulandığında, tüm modülü özelliklerini etkileyen bir sertleşme neden elastik özelliklerini değiştirir. Hareket ve sertlik performansları bir hava kompresörü ve odaları ve sertleştirme kanalında vakum etkinleştirmek için bir vakum pompası harekete geçirmek için üç basınç valfleri dahil harici kurulum tarafından kontrol edilir. Sezgisel bir kullanıcı arayüzü modülü içindeki tahrik ve vakum basınçları kontrol sağlar. Bu yazıda fabricatio detaylarıBu manipülatör ve raporlar temel hareket yetenekleri en önemli sonuçları tek modül n süreci. Cihazın modüler doğası göz önüne alındığında, imalat ve sadece tek bir modülün performans değerlendirmesi de sonuçları uzatılabilir ve iki veya daha fazla modül entegre bir çok modül manipülatör temel davranışlarını tahmin sağlar.

<table border="0">
	<tbody>
		<tr>
			<td>Ecoflex 00-50 Trial Kit</td>
			<td>SmoothOn</td>
			<td></td>
			<td>Used for the fabrication of the soft unit, combining equal amounts of liquid parts A (the base) and B (the catalyst)</td>
		</tr>
		<tr>
			<td>Latex</td>
			<td>Antichit&#224; Belsito</td>
			<td></td>
			<td>Used for the fabrication of the granular jamming membrane</td>
		</tr>
		<tr>
			<td>Peroxide-Cured Silicone Tubing</td>
			<td>Cole Parmer</td>
			<td>T-06411-59</td>
			<td>Used for actuating the chambers and applying vacuum</td>
		</tr>
		<tr>
			<td>PET expandable braided sleeving</td>
			<td>RS</td>
			<td>408-249</td>
			<td>Used for the fabrication of the external braided sheath</td>
		</tr>
		<tr>
			<td>Silicone Rubber</td>
			<td>Momentive</td>
			<td>127374</td>
			<td>Used to fix the actuation tubes to the module</td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td>Cole Parmer</td>
			<td>EW-06720-40</td>
			<td>Used to fix the latex membrane to the vacuum tube</td>
		</tr>
		<tr>
			<td>Fume hood Secuflow</td>
			<td>Groupe Waldner</td>
			<td></td>
			<td>Working space</td>
		</tr>
		<tr>
			<td>Precision scale</td>
			<td>KERN EW</td>
			<td></td>
			<td>Used to weight silicone, latex and coffee powder</td>
		</tr>
		<tr>
			<td>Oven/degasser</td>
			<td>Heraeus</td>
			<td></td>
			<td>Used to degass the silicone and reduce its cure time</td>
		</tr>
		<tr>
			<td>Vacuum pump</td>
			<td>DVP Vacuum Technology</td>
			<td></td>
			<td>Used to apply vacuum to the latex membrane</td>
		</tr>
	</tbody>
</table>

design and fabrication of an elastomeric unit for soft modular robots in minimally invasive surgery

Son yıllarda, yumuşak robotik teknolojileri nedeniyle yapılandırılmamış ortamlarda kendi kendinden güvenli etkileşime tıp alanında artan ilgi uyandırmıştır. Aynı zamanda, yeni yöntem ve teknikler cerrahi operasyonlar invazivliğini azaltmak için geliştirilmiştir. Minimal İnvazif Cerrahi (MIS) başarıyla karın müdahaleler için istihdam edilmiştir, ancak standart MIS prosedürler esas klinisyenin becerisi sınırlayan sert veya yarı sert araçları dayanmaktadır. Bu kağıt MIS için yumuşak ve yüksek hünerli manipülatör sunar. Manipülatör ahtapot kolunun biyolojik yetenekleri esinlenerek ve modüler bir yaklaşımla tasarlanmıştır. Her bir modül daha modül entegre edildiğinde böylece yüksek maharet ve çok yönlülüğü elde, aynı işlevsel özelliklere sunuyor. Kağıt castin tarafından imal edilmektedir tek bir birim, gelişimi için gerekli tasarım, imalat süreci ve malzemeleri ayrıntılarıBelirli bir kalıp içine g silikon. Sonuç uzaması ve ünitenin yönlü eğilmesine olanak üç esnek pnömatik aktüatörler içeren bir elastomerik silindir içinde oluşur. Bir dış kılıf örgülü modülünün hareketini artırır. Her bir modülün merkezinde bir zerrecik halinde sıkışma tabanlı bir mekanizma, görevleri sırasında, yapının sertliğini değişir. Testler modülü 120 ° &#39;ye kadar eğmek ve başlangıç ​​uzunluğunun% 66 kadar uzatmak mümkün olduğunu göstermektedir. Modül 47 N maksimum güç üretir ve onun sertliği% 36 kadar artırabilir.

Protokolde tanımlanan imalat, çeşitli fazlar Şekil 3 &#39;de gösterilmiştir. Tekniğin etkililiğini ve son prototip sonuçlarını değerlendirmek için, modül farklı çalışma koşullarında test edilmiştir. Harici kurulum harekete ve modülün sertlik hem kontrolünü sağlar. Üç vana aktive bir hava kompresör içerir. Bu odalar entegre siliconic tüplere bağlanmış ve basınçlanması sağlandı edilir. Bir vakum pompası modülü sertliği kontrolü için...

Watch this Scientific Journal Video about Design and Fabrication of an Elastomeric Unit for Soft Modular Robots in Minimally Invasive Surgery at JoVE.com

Design and Fabrication of an Elastomeric Unit for Soft Modular Robots in Minimally Invasive Surgery

This paper describes the design and fabrication of a soft unit for surgical manipulators. The base module includes three flexible fluidic actuators to achieve omnidirectional bending and elongation, and a granular jamming-based mechanism to enable stiffness control. A complete mechanical characterization is also reported.

Tasarım ve Yumuşak Modüler Robotlar için bir Elastomerik Birimi Fabrikasyon Minimal İnvaziv Cerrahi

In recent years, soft robotics technologies have aroused increasing interest in the medical field due to their intrinsically safe interaction in ...

design-fabrication-an-elastomeric-unit-for-soft-modular-robots

Research

JoVE Journal

Bioengineering

8.9K Görüntüleme.  Scuola Superiore Sant'Anna. This paper describes the design and fabrication of a soft unit for surgical manipulators. The base module includes three flexible fluidic actuators to achieve omnidirectional bending and elongation, and a granular jamming-based mechanism to enable stiffness control. A complete mechanical characterization is also reported. 

Video: Design and Fabrication of an Elastomeric Unit for Soft Modular Robots in Minimally Invasive Surgery

Elastomeric PGS Scaffolds in Arterial Tissue Engineering

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and increasing obesity1. Currently, bypass surgery using autologous vessels, allografts, and synthetic grafts are known as a commonly used for arterial substitutes2. However, these grafts have limited applications when an inner diameter of arteries is less than 6 mm due to low availability, thrombotic complications, compliance mismatch, and late intimal hyperplasia3,4. To overcome these limitations, tissue engineering has been successfully applied as a promising alternative to develop small-diameter arterial constructs that are nonthrombogenic, robust, and compliant. Several previous studies have developed small-diameter arterial constructs with tri-lamellar structure, excellent mechanical properties and burst pressure comparable to native arteries5,6. While high tensile strength and burst pressure by increasing collagen production from a rigid material or cell sheet scaffold, these constructs still had low elastin production and compliance, which is a major problem to cause graft failure after implantation. Considering these issues, we hypothesized that an elastometric biomaterial combined with mechanical conditioning would provide elasticity and conduct mechanical signals more efficiently to vascular cells, which increase extracellular matrix production and support cellular orientation.
The objective of this report is to introduce a fabrication technique of porous tubular scaffolds and a dynamic mechanical conditioning for applying them to arterial tissue engineering. We used a biodegradable elastomer, poly (glycerol sebacate) (PGS)7 for fabricating porous tubular scaffolds from the salt fusion method. Adult primary baboon smooth muscle cells (SMCs) were seeded on the lumen of scaffolds, which cultured in our designed pulsatile flow bioreactor for 3 weeks. PGS scaffolds had consistent thickness and randomly distributed macro- and micro-pores. Mechanical conditioning from pulsatile flow bioreactor supported SMC orientation and enhanced ECM production in scaffolds. These results suggest that elastomeric scaffolds and mechanical conditioning of bioreactor culture may be a promising method for arterial tissue engineering.

Elastomeric PGS scaffolds with vascular smooth muscle cells cultured in a pulsatile flow bioreactor may lead to promising small-diameter arterial constructs with native ECM production in a relatively short culture period.

Elastomerik PGS Arter Doku Mühendisliği Yapı iskeleleri

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and ...

Biyomühendislik

Polymeric Microneedle Array Fabrication by Photolithography

This manuscript describes the fabrication of polymeric microneedle (MN) arrays by photolithography. It involves a simple mold-free process by using a photomask consisting of embedded micro-lenses. Embedded micro-lenses were found to influence MN geometry (sharpness). Robust MN arrays with tip diameters ranging between 41.5 &#181;m &#177; 8.4 &#181;m and 71.6 &#181;m &#177; 13.7 &#181;m, with two different lengths (1,336 &#181;m &#177; 193 &#181;m and 957 &#181;m &#177; 171 &#181;m) were fabricated. These MN arrays may provide potential applications in delivery of low molecular and macromolecular therapeutic agents through skin.

Here, we present a protocol describing a mold-free fabrication process of the polymeric microneedles by photolithography.

Fotolitografiyle polimerik Microneedle Dizi Fabrikasyon

This manuscript describes the fabrication of polymeric microneedle (MN) arrays by photolithography. It involves a simple mold-free process by using a ...

Rod-based Fabrication of Customizable Soft Robotic Pneumatic Gripper Devices for Delicate Tissue Manipulation

Soft compliant gripping is essential in delicate surgical manipulation for minimizing the risk of tissue grip damage caused by high stress concentrations at the point of contact. It can be achieved by complementing traditional rigid grippers with soft robotic pneumatic gripper devices. This manuscript describes a rod-based approach that combined both 3D-printing and a modified soft lithography technique to fabricate the soft pneumatic gripper. In brief, the pneumatic featureless mold with chamber component is 3D-printed and the rods were used to create the pneumatic channels that connect to the chamber. This protocol eliminates the risk of channels occluding during the sealing process and the need for external air source or related control circuit. The soft gripper consists of a chamber filled with air, and one or more gripper arms with a pneumatic channel in each arm connected to the chamber. The pneumatic channel is positioned close to the outer wall to create different stiffness in the gripper arm. Upon compression of the chamber which generates pressure on the pneumatic channel, the gripper arm will bend inward to form a close grip posture because the outer wall area is more compliant. The soft gripper can be inserted into a 3D-printed handling tool with two different control modes for chamber compression: manual gripper mode with a movable piston, and robotic gripper mode with a linear actuator. The double-arm gripper with two actuatable arms was able to pick up objects of sizes up to 2 mm and yet generate lower compressive forces as compared to elastomer-coated and non-coated rigid grippers. The feasibility of having other designs, such as single-arm or hook gripper, was also demonstrated, which further highlighted the customizability of the soft gripper device, and it's potential to be used in delicate surgical manipulation to reduce the risk of tissue grip damage.

This protocol describes a rod-based approach, combining 3D-printing and soft lithography techniques for fabricating the soft gripper devices. This approach eliminates the need for an external air source by incorporating a chamber component and reduces the chance of occlusion during the sealing process, particularly for miniaturized pneumatic channels.

Narin Doku Manipülasyon için Özelleştirilebilir Yumuşak Robotik Pnömatik Tutucu Cihazlar Rod tabanlı Fabrikasyon

Soft compliant gripping is essential in delicate surgical manipulation for minimizing the risk of tissue grip damage caused by high stress concentrations ...

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; ...

Biomechanical Characterization of Human Soft Tissues Using Indentation and Tensile Testing

Regenerative medicine aims to engineer materials to replace or restore damaged or diseased organs. The mechanical properties of such materials should mimic the human tissues they are aiming to replace; to provide the required anatomical shape, the materials must be able to sustain the mechanical forces they will experience when implanted at the defect site. Although the mechanical properties of tissue-engineered scaffolds are of great importance, many human tissues that undergo restoration with engineered materials have not been fully biomechanically characterized. Several compressive and tensile protocols are reported for evaluating materials, but with large variability it is difficult to compare results between studies. Further complicating the studies is the often destructive nature of mechanical testing. Whilst an understanding of tissue failure is important, it is also important to have knowledge of the elastic and viscoelastic properties under more physiological loading conditions.
This report aims to provide a minimally destructive protocol to evaluate the compressive and tensile properties of human soft tissues. As examples of this technique, the tensile testing of skin and the compressive testing of cartilage are described. These protocols can also be directly applied to synthetic materials to ensure that the mechanical properties are similar to the native tissue. Protocols to assess the mechanical properties of human native tissue will allow a benchmark by which to create suitable tissue-engineered substitutes.

Tissue biomechanics is important for maintaining cell shape and function and for determining phenotype. This report demonstrates non-destructive mechanical protocols for characterizing elastic and viscoelastic properties of human soft tissues, which can be directly applied to tissue-engineered substrates to allow a close matching of engineered materials to native tissue.

Girinti ve Çekme Test Kullanarak İnsan Yumuşak Doku biyomekanik Karakterizasyonu

Regenerative medicine aims to engineer materials to replace or restore damaged or diseased organs. The mechanical properties of such materials should ...

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and dampen loads in the spine. The IVD consists of a proteoglycan-rich nucleus pulposus (NP) and a collagen-rich annulus fibrosis (AF) sandwiched by cartilaginous end-plates. Together with the adjacent vertebrae, the vertebrae-IVD structure forms a functional spine unit (FSU). These microstructures contain unique cell types as well as unique extracellular matrices. Whole organ culture of the FSU preserves the native extracellular matrix, cell differentiation phenotypes, and cellular-matrix interactions. Thus, organ culture techniques are particularly useful for investigating the complex biological mechanisms of the IVD. Here, we describe a high-throughput approach for culturing whole lumbar mouse FSUs that provides an ideal platform for studying disease mechanisms and therapies for the IVD. Furthermore, we describe several applications that utilize this organ culture method to conduct further studies including contrast-enhanced microCT imaging and three-dimensional high-resolution finite element modeling of the IVD.

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

Whole organ culture of the intervertebral disc (IVD) preserves the native extracellular matrix, cell phenotypes, and cellular-matrix interactions. Here we describe an IVD culture system using mouse lumbar and caudal IVDs in their functional spinal units and several applications utilizing this system.

bir<em&gt; İn Vitro</emKemirgen İntervertebral Disk&gt; Organ Kültürü Modeli

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and ...

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver engineering, including in vivo organ decellularization followed by repopulation, has emerged as a promising approach over ex vivo liver engineering. However, postoperative survival was not achieved. The aim of this study is to develop a novel surgical technique of in vivo selective liver lobe perfusion in rats as a prerequisite for in vivo liver engineering. We generate a circuit bypass only through the left lateral lobe. Then, the left lateral lobe is perfused with heparinized saline. The experiment is performed with 4 groups (n = 3 rats per group) based on different perfusion times of 20 min, 2 h, 3 h, and 4 h. Survival, as well as the macroscopically visible change of color and the histologically determined absence of blood cells in the portal triad and the sinusoids, is taken as an indicator for a successful model establishment. After selective perfusion of the left lateral lobe, we observe that the left lateral lobe, indeed, turned from red to faint yellow. In a histological assessment, no blood cells are visible in the branch of the portal vein, the central vein, and the sinusoids. The left lateral lobe turns red after reopening the blocked vessels. 12/12 rats survived the procedure for more than one week. We are the first to report a surgical model for in vivo single liver lobe perfusion with a long survival period of more than one week. In contrast to the previously published report, the most important advantage of the technique presented here is that perfusion of 70% of the liver is maintained throughout the whole procedure. The establishment of this technique provides a foundation for in vivo partial liver engineering in rats, including decellularization and recellularization.

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

We establish a novel surgical technique for an in vivo single liver lobe perfusion model in rat as a prerequisite for further studying in vivo partial liver engineering in the future.

Sıçan Vivo içinde kısmi karaciğer Dairesi'nde için bir temel olarak yeni bir cerrahi teknik

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver ...

Bioinspired Soft Robot with Incorporated Microelectrodes

Bioinspired soft robotic systems that mimic living organisms using engineered muscle tissue and biomaterials are revolutionizing the current biorobotics paradigm, especially in biomedical research. Recreating artificial life-like actuation dynamics is crucial for a soft-robotic system. However, the precise control and tuning of actuation behavior still represents one of the main challenges of modern soft robotic systems. This method describes a low-cost, highly scalable, and easy-to-use procedure to fabricate an electrically controllable soft robot with life-like movements that is activated and controlled by the contraction of cardiac muscle tissue on a micropatterned sting ray-like hydrogel scaffold. The use of soft photolithography methods makes it possible to successfully integrate multiple components in the soft robotic system, including micropatterned hydrogel-based scaffolds with carbon nanotubes (CNTs) embedded gelatin methacryloyl (CNT-GelMA), poly(ethylene glycol) diacrylate (PEGDA), flexible gold (Au) microelectrodes, and cardiac muscle tissue. In particular, the hydrogels alignment and micropattern are designed to mimic the muscle and cartilage structure of the sting ray. The electrically conductive CNT-GelMA hydrogel acts as a cell scaffold that improves the maturation and contraction behavior of cardiomyocytes, while the mechanically robust PEGDA hydrogel provides structural cartilage-like support to the whole soft robot. To overcome the hard and brittle nature of metal-based microelectrodes, we designed a serpentine pattern that has high flexibility and can avoid hampering the beating dynamics of cardiomyocytes. The incorporated flexible Au microelectrodes provide electrical stimulation across the soft robot, making it easier to control the contraction behavior of cardiac tissue.

A bioinspired scaffold is fabricated by a soft photolithography technique using mechanically robust and electrically conductive hydrogels. The micropatterned hydrogels provide directional cardiomyocyte cell alignment, resulting in a tailored direction of actuation. Flexible microelectrodes are also integrated into the scaffold to bring electrical controllability for a self-actuating cardiac tissue.

Anonim Mikroelektrotlu Bioinspired Yumuşak Robot

Bioinspired soft robotic systems that mimic living organisms using engineered muscle tissue and biomaterials are revolutionizing the current biorobotics ...

Simple, Affordable, and Modular Patterning of Cells using DNA

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells of the same or different type, micropatterning techniques have proved useful. DNA Programmed Assembly of Cells (DPAC) is a micropatterning technique that targets the adhesion of cells to a substrate or other cells using DNA hybridization. The most basic operations in DPAC begin with decorating cell membranes with lipid-modified oligonucleotides, then flowing them over a substrate that has been patterned with complementary DNA sequences. Cells adhere selectively to the substrate only where they find a complementary DNA sequence. Non-adherent cells are washed away, revealing a pattern of adherent cells. Additional operations include further rounds of cell-substrate or cell-cell adhesion, as well as transferring the patterns formed by DPAC to an embedding hydrogel for long-term culture. Previously, methods for patterning oligonucleotides on surfaces and decorating cells with DNA sequences required specialized equipment and custom DNA synthesis, respectively. We report an updated version of the protocol, utilizing an inexpensive benchtop photolithography setup and commercially available cholesterol modified oligonucleotides (CMOs) deployed using a modular format. CMO-labeled cells adhere with high efficiency to DNA-patterned substrates. This approach can be used to pattern multiple cell types at once with high precision and to create arrays of microtissues embedded within an extracellular matrix. Advantages of this method include its high resolution, ability to embed cells into a three-dimensional microenvironment without disrupting the micropattern, and flexibility in patterning any cell type.

Here we present a protocol to micropattern cells at single-cell resolution using DNA-programmed adhesion. This protocol uses a benchtop photolithography platform to create patterns of DNA oligonucleotides on a glass slide and then labels cell membranes with commercially available complementary oligonucleotides. Hybridization of the oligos results in programmed cell adhesion.

DNA Kullanarak Hücrelerin Basit, Uygun Fiyatlı ve Modüler Deseni

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells ...

Voxel Printing Anatomy: Design and Fabrication of Realistic, Presurgical Planning Models through Bitmap Printing

Most applications of 3-dimensional (3D) printing for presurgical planning have been limited to bony structures and simple morphological descriptions of complex organs due to the fundamental limitations in accuracy, quality, and efficiency of the current modeling paradigm. This has largely ignored the soft tissue critical to most surgical specialties where the interior of an object matters and anatomical boundaries transition gradually. Therefore, the needs of the biomedical industry to replicate human tissue, which displays multiple scales of organization and varying material distributions, necessitate new forms of representation. 
Presented here is a novel technique to create 3D models directly from medical images, which are superior in spatial and contrast resolution to current 3D modeling methods and contain previously unachievable spatial fidelity and soft tissue differentiation. Also presented are empirical measurements of novel, additively manufactured composites that span the gamut of material stiffnesses seen in soft biological tissues from MRI and CT. These unique volumetric design and printing methods allow for deterministic and continuous adjustment of material stiffness and color. This capability enables an entirely new application of additive manufacturing to presurgical planning: mechanical realism. As a natural complement to existing models that provide appearance matching, these new models also allow medical professionals to "feel" the spatially varying material properties of a tissue simulant-a critical addition to a field in which tactile sensation plays a key role.

This method demonstrates a voxel-based 3D printing workflow, which prints directly from medical images with exact spatial fidelity and spatial/contrast resolution. This enables the precise, graduated control of material distributions through morphologically complex, graduated materials correlated to radiodensity without loss or alteration of data.

Voxel Baskı Anatomisi: Bitmap Baskı Ile Gerçekçi, Presurgical Planlama Modellerinin Tasarımı ve İmalatı

Most applications of 3-dimensional (3D) printing for presurgical planning have been limited to bony structures and simple morphological descriptions of ...