이 연구는 재료의 연구와 교육에 대한 국립 과학 재단 (National Science Foundation) 협력 (PREM) 부여 번호 DMR - 0934218에 의해 지원되었다. 또한 연구 자원에 대한 국립 센터에서 보너스 번호 2G12RR013646 - 11에 의해 지원되었다. 내용은 전적으로 저자의 책임이며 반드시 연구 자원이나 국립 보건원에 대한 국립 센터의 공식 견해를 대변하지 않습니다.

1. 실버 종자 준비 <ol><li> 임의의 질량을 감수하면서 deionized (DI) 물 10 ML로 혼합하여 실버 질산 (AgNO 3) 재고 솔루션을 준비합니다. 솔루션의 molarity을 계산합니다. 라이트에서 분리 어두운 장소에 솔루션을 보관하십시오. </li><li> 5 MM 솔루션을 만들기 위해 DI 물 10 ML에 나트륨 구연산 tribasic의 14.7 MG (나 3 C 6 H 5 O 7) 추가합니다. 파우더가 해산 때까지 병을 흔들어. </li><li> 40 MM 솔루션을 만들기 위해 DI 물 10 ML 다른 유리병에 나트륨 borohydride (NaBH 4) 15.1 밀리그램을 추가합니다. 즉시 병을를 닫습니다. 부드럽게 손으로 솔루션을 흔들 얼음 비커에 넣습니다. 냉장고에 비커를 놓고 (T1 = 0) 타이머를 시작합니다. 갓 만들어진 솔루션은 충분히 그것을 아래로 냉각 시간 15 분 사용됩니다. </li><li> 실버 질산, 1.1)의 주식 솔루션에서, 0.25 MM 10 ML을 준비합니다. 유리병와 ST의 감동 자석 장소예술 감동. </li><li> 1.4 나트륨 구연산 tribasic 솔루션 1.2))의 0.25 ML을 추가합니다. </li><li> t 1 = 15 분 냉장고에서 나트륨 borohydride 솔루션, 1.3)를 제거하십시오. 피펫이 솔루션 0.4 ML을 가지고) 1.5에 추가 사용. 참고 : 단일 빠른 스트로크에 솔루션을 추가합니다. 색깔이 노란색으로 바뀝니다. 5 분 솔루션을 저어. </li><li> t 1 = 20 분 중지 감동, 유리병에서 자석을 제거하고 어두운 장소에 보관. 약병을 닫지 마십시오. </li><li> 사용하기 전에 최소한 2 시간 동안 실온에서 어둠의 솔루션을 유지. 가급적 준비 한 주일 이내에 씨앗을 사용합니다. </li></ol> 2. 성장 솔루션 준비 <ol><li> DI 워터 10 ML에 140 밀리그램을 추가하여 아스코르비 산 (C 6 H 8 O 6)의 80 MM을 준비합니다. </li><li> 골드 염화 (HAuCl 4) 농축 용액 10 ML을 준비합니다. 솔루션의 molarity을 계산합니다. 솔루션을 유지n은 빛으로부터 격리. </li><li> DI 워터 20 ML과 유리병에 364 MG를 추가하여 - cetyltrimethylammonium의 브로마이드 50 MM (C 19 H 42 BrN CTAB)의 20 ML을 준비합니다. 즉시 유리병에 감동 자석을 배치하고 30 따뜻한 접시에 감동을 시작 ° C. CTAB 분말이 완전히 해산 및 솔루션은 투명 판의 히터 해제하지만, 단계 2.7을 통해 감동 유지)되는 후. </li><li> 4.9x10 -2 mm의 최종 molarity를 ​​구하는 해결책 2.3)에 솔루션 1.1) 추가합니다. (T 2 = 0) 타이머를 시작합니다. </li><li> t 2 = 1 분, 0.25 mm의 최종 molarity를 ​​얻을 2.4)에 솔루션 2.2) 추가합니다. </li><li> t 2 = 2 분) 2.5-2.1의 0.1 ML)를 추가합니다. 이 솔루션은 무색으로 바뀔 것이다. </li><li> t 2 = 2 분 20 초)이 2.6-1.8의 0.05 ML) (실버 씨앗)을 추가합니다. 15 분에 대한 정지를 저어. 정지는 처음에 파란색 후 갈색으로 바뀔 것이다. </li><li> t 2 = 17 분 정지의 감동, m을 제거agnet 24 시간 동안 실온에서 정학을 유지. </li></ol> 3. 이미징, 특성 또는 실험에 대해 CTAB에서 골드 nanostars 분리 참고 : CTAB는 실온에서 구체화 수 있습니다. 30 ° C로 골드 콜로이드 최대 크리스탈 열을 분해 또는 수정이 용해 때까지 탭 물을 고온에 병을 담가합니다. <ol><li> 2 분 정지를 Sonicate. </li><li> 730 rcf에서 5 분 현탁액을 원심. Nanostars은 튜브의 벽에 축적됩니다. </li><li> nanostars을 제거하지 않도록 관리하는 피펫와 마찬가지로 많은 정지 제거합니다. </li><li> 튜브에 DI 물을 추가 2 분 sonicate. </li><li> 460 rcf에서 3 분 현탁액을 원심. 정지 덜 CTAB를 포함, 따라서 낮은 원심력은 nanostars을 구분이 필요합니다. </li><li> ) 단계 3.3) 및 3.4를 반복합니다. </li><li> 380 rcf에서 3 분 서스펜션과 원심에 DI 워터를 추가합니다. </li><li> 반복 하오<html> t 3.3) 및 3.4) 단계를 반복합니다. nanostars는 이미징, 분광, 또는 실험을위한 준비가되어 있습니다. </li></ol> 4. 대표 결과 : 그림 1은 실버 씨앗 JEOL 2010 - F TEM을 사용하여 몇 군데의 전송 전자 현미경 (TEM) 이미지를 보여줍니다. 씨앗은 구형 모양과 15 nm의의 평균 크기를했습니다. 골드 nanostars은 히타치를 사용하여 몇 군데 아르 S - 5500 전자 현미경 (SEM) 모드를 스캔 인치 그림 2는 우리의 방법과 합성 nanostars의 증가 배율을 보여줍니다. 스타 모양의 입자 콜로이드의 모든 입자의 약 70 %입니다. 비 형성 스타 decahedra 및 rhomboids의 비정질 클러스터 (표시되지 않음)처럼 나타납니다. 그림 3은 여러 싱글 골드 nanostars을 보여줍니다. 200 NM에서 300 NM 및 도움말의 개수에 nanostars 범위의 크기는 7에서 10에 따라 다릅니다. 황금 그들이 합성 후 최소 1 개월 그 모양을 그대로 유지이 방법 CTAB에 남아 의해 합성 nanoparticles 경우. e_content &quot;&gt; 우리는 Olis 캐리 - 14 분광 광도계를 사용하여 실버 종자와 nanostars의 흡수 스펙트럼을 측정. 종자의 최대 흡수가 400 NM에서 되었음 nanostars의 최대 흡수 800 NM 850 NM (그림 4의 사이 동안 ). <img src="/files/ftp_upload/3570/3570fig1.jpg" alt="그림 1" /> 그림 1. 은빛 씨앗의 전송 전자 현미경 이미지. <img src="/files/ftp_upload/3570/3570fig2.jpg" alt="그림 2" /> 그림 2. 금을 nanostars의 전자 현미경 이미지를 검사합니다. <img src="/files/ftp_upload/3570/3570fig3.jpg" alt="그림 3" /> 그림 3. 단일 골드 nanostars의 전자 현미경 이미지를 검사합니다. <img src="/files/ftp_upload/3570/3570fig4.jpg" alt="그림 6" /> 그림 4. 표준화 흡수 실버 종자의 스펙트럼 (점선)과 금nanostars (실선).

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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Seed-mediated+growth+approach+for+shape-controlled+synthesis+of+spheroidal+and+rod-like+gold+nanoparticles+using+a+surfactant+template.">Seed-mediated growth approach for shape-controlled synthesis of spheroidal and rod-like gold nanoparticles using a surfactant template.</a> Advanced Materials. 13, 1389-1393 (2001).</li><li>Xiao, J., Qi, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surfactant-assisted,+shape-controlled+synthesis+of+gold+nanocrystals.">Surfactant-assisted, shape-controlled synthesis of gold nanocrystals.</a> Nanoscale. 3, 1383-1396 (2011).</li><li>Tao, A. R., Habas, S., Yang, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shape+control+of+colloidal+metal+nanocrystals.">Shape control of colloidal metal nanocrystals.</a> Small. 4, 310-325 (2008).</li><li>Cole, J. R., Mirin, N. A., Knight, M. W., Goodrich, G. P., Halas, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photothermal+Efficiencies+of+Nanoshells+and+Nanorods+for+Clinical+Therapeutic+Applications.">Photothermal Efficiencies of Nanoshells and Nanorods for Clinical Therapeutic Applications.</a> Journal of Physical Chemistry C. 113, 12090-12094 (2009).</li><li>Choi, J. S., Park, J. C., Nah, H., Woo, S., Oh, J., Kim, K. M., Cheon, G. J., Chang, Y., Yoo, J., Cheon, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+hybrid+nanoparticle+probe+for+dual-modality+positron+emission+tomography+and+magnetic+resonance+imaging.">A hybrid nanoparticle probe for dual-modality positron emission tomography and magnetic resonance imaging.</a> Angew. Chem. Int. Ed. Engl. 47, 6259-6262 (2008).</li><li>Chithrani, B. D., Ghazani, A. A., Chan, W. C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determining+the+Size+and+Shape+Dependence+of+Gold+Nanoparticle+Uptake+into+Mammalian+Cells.">Determining the Size and Shape Dependence of Gold Nanoparticle Uptake into Mammalian Cells.</a> Nano Letters. 6, 662-668 (2006).</li></ol>

관심 없음 충돌 선언하지 않습니다.

이 작품에서 우리는 실버 종자를 사용하여 골드 nanostars를 종합하는 방법을 제시합니다. 우리는 실버 씨앗이 nanostars의 70 %를 생산 수율 결과 것으로 나타났습니다. nanostars 800 NM 850 NM 7, 23 사이를 중심으로 그들의 표면 plasmon 공진 모드에 해당하는 가까운 적외선 흡수 피크를,있다. 이러한 속성 속성은 우리의 골드 nanostars 이러한 사진 열 절제로 24-26 생물 의학 응용 프로그램에 사용...

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 순도 </td></tr><tr><td> 나트륨 구연산 tribasic의 탈수 </td><td> 시그마 </td><td> S4641 </td><td> 99.0 % </td></tr><tr><td> 실버 질산염 </td><td> 올드 리치 </td><td> 204,390 </td><td> 99.9999 % </td></tr><tr><td> 나트륨 borohydride </td><td> 올드 리치 </td><td> 213,462 </td><td> 99% </td></tr><tr><td> L - 아스코르비 산 </td><td> 시그마 - 올드 리치 </td><td> 255,564 </td><td> 99 이상 % </td></tr><tr><td> 골드 염화물 trihydrate </td><td> 올드 리치 </td><td> 520,918 </td><td> 99.9 이상 % </td></tr><tr><td> Hexadecyltrimethylammonium의 브로마이드 (CTAB) </td><td> 시그마 </td><td> H6269 </td><td></td></tr></tbody></table><table border="1"><tbody><tr><td> 장비의 명칭 </td><td> 회사 </st룽&gt; </td><td> 댓글 </td></tr><tr><td> JEOL 2010 - F </td><td> JEOL </td><td> 전송 전자 현미경 </td></tr><tr><td> 히타치 S - 5500 </td><td> 히타치 </td><td> 전자 현미경 모드를 스캔에 사용 </td></tr><tr><td> Olis 캐리 - 14 분광 광도계 </td><td> Olis </td><td> 분광 광도계 </td></tr></tbody></table>

gold nanostar synthesis with a silver seed mediated growth method

나노 스케일 colloids의 물리적, 화학적 및 광학 성질들은 재료 성분, 크기와 모양 1-5에 따라 달라집니다. 사진 열 절제, 약물 전달 및 기타 여러 생명 의학 애플 리케이션 6 나노 colloids를 사용하여 큰 관심이 있습니다. 골드는 특히 때문에 낮은 독성 7-9 중 사용됩니다. 금속 나노 colloids의 속성들은 강력한 표면 plasmon 공명 10 가질 수 있습니다. 표면 plasmon 공명 모드의 절정은 금속 나노 colloids의 구조와 구성에 따라 달라집니다. 표면 plasmon 공명 모드가 빛을 자극되기 때문에 생물 학적 조직 투과율이 11, 12 최대한의 어디에 가까운 적외선의 최대 흡광도를 가질 필요가있다. 우리는 또한 별 모양 nanoparticles 13-15 또는 nanostars 16 알려져 별 모양의 금 콜로이드를 합성하는 방법을 제시한다. 이 방법은로 기반황금 colloids 17-22의 비등 방성 성장을위한 nucleating 에이전트로 사용되는 은색 씨앗을 포함 olution. 결과 금 콜로이드의 스캐닝 전자 현미경 (SEM) 분석은 nanostructures의 70 %가 nanostars 것을 보여주었다. 입자의 다른 30 ​​% decahedra 및 rhomboids의 비정질 클러스터했다. nanostars의 흡광도 피크는 가까운 적외선 (840 NM)에 있어야 발견했습니다. 따라서, 우리의 방법은 특히 사진 열 절제를 위해, 생명 의학 애플 리케이션에 적합한 골드 nanostars을 생산하고 있습니다.

Watch this Scientific Journal Video about Gold Nanostar Synthesis with a Silver Seed Mediated Growth Method at JoVE.com

Gold Nanostar Synthesis with a Silver Seed Mediated Growth Method

우리는 은색 씨앗 중재의 성장 방법을 사용하여 별 모양의 골드 nanostars을 합성. nanostars의 직경이 200-300 nm의 범위에서 및 도움말의 숫자 7에서 10에 따라 다릅니다. nanoparticles는 가까운 적외선 중심으로 광범위한 표면 plasmon 공진 모드를있다.

실버 씨앗 중재 성장 방법과 골드 Nanostar 합성

The physical, chemical and optical properties of nano-scale colloids depend on their material composition, size and shape 1-5. There is a great interest ...

gold-nanostar-synthesis-with-a-silver-seed-mediated-growth-method

Research

JoVE Journal

Bioengineering

24.9K 조회수.  The University of Texas at San Antonio. 우리는 은색 씨앗 중재의 성장 방법을 사용하여 별 모양의 골드 nanostars을 합성. nanostars의 직경이 200-300 nm의 범위에서 및 도움말의 숫자 7에서 10에 따라 다릅니다. nanoparticles는 가까운 적외선 중심으로 광범위한 표면 plasmon 공진 모드를있다.

비디오: Gold Nanostar Synthesis with a Silver Seed Mediated Growth Method

Axon Stretch Growth: The Mechanotransduction of Neuronal Growth

During pre-synaptic embryonic development, neuronal processes traverse short distances to reach their targets via growth cone. Over time, neuronal somata are separated from their axon terminals due to skeletal growth of the enlarging organism (Weiss 1941; Gray, Hukkanen et al. 1992). This mechanotransduction induces a secondary mode of neuronal growth capable of accommodating continual elongation of the axon (Bray 1984; Heidemann and Buxbaum 1994; Heidemann, Lamoureux et al. 1995; Pfister, Iwata et al. 2004).
Axon Stretch Growth (ASG) is conceivably a central factor in the maturation of short embryonic processes into the long nerves and white matter tracts characteristic of the adult nervous system. To study ASG in vitro, we engineered bioreactors to apply tension to the short axonal processes of neuronal cultures (Loverde, Ozoka et al. 2011). Here, we detail the methods we use to prepare bioreactors and conduct ASG. First, within each stretching lane of the bioreactor, neurons are plated upon a micro-manipulated towing substrate. Next, neurons regenerate their axonal processes, via growth cone extension, onto a stationary substrate. Finally, stretch growth is performed by towing the plated cell bodies away from the axon terminals adhered to the stationary substrate; recapitulating skeletal growth after growth cone extension.
Previous work has shown that ASG of embryonic rat dorsal root ganglia neurons are capable of unprecedented growth rates up to 10mm/day, reaching lengths of up to 10cm; while concurrently resulting in increased axonal diameters (Smith, Wolf et al. 2001; Pfister, Iwata et al. 2004; Pfister, Bonislawski et al. 2006; Pfister, Iwata et al. 2006; Smith 2009). This is in dramatic contrast to regenerative growth cone extension (in absence of mechanical stimuli) where growth rates average 1mm/day with successful regeneration limited to lengths of less than 3cm (Fu and Gordon 1997; Pfister, Gordon et al. 2011). Accordingly, further study of ASG may help to reveal dysregulated growth mechanisms that limit regeneration in the absence of mechanical stimuli.

A unique tissue engineering method was developed to elongate numerous nerve fibers in culture by recapitulating axon stretch growth; a form of nervous system growth whereby nerves elongate in conjunction with growth of the enlarging body.

액슨 스트레치 성장의 연결을 성장의 Mechanotransduction

During pre-synaptic embryonic development, neuronal processes traverse short distances to reach their targets via growth cone. Over time, neuronal somata ...

연구

생체공학

Synthesis, Assembly, and Characterization of Monolayer Protected Gold Nanoparticle Films for Protein Monolayer Electrochemistry

Colloidal gold nanoparticles protected with alkanethiolate ligands called monolayer protected gold clusters (MPCs) are synthesized and subsequently incorporated into film assemblies that serve as adsorption platforms for protein monolayer electrochemistry (PME). PME is utilized as the model system for studying electrochemical properties of redox proteins by confining them to an adsorption platform at a modified electrode, which also serves as a redox partner for electron transfer (ET) reactions. Studies have shown that gold nanoparticle film assemblies of this nature provide for a more homogeneous protein adsorption environment and promote ET without distance dependence compared to the more traditional systems modified with alkanethiol self-assembled monolayers (SAM).1-3 In this paper, MPCs functionalized with hexanethiolate ligands are synthesized using a modified Brust reaction4 and characterized with ultraviolet visible (UV-Vis) spectroscopy, transmission electron microscopy (TEM), and proton (1H) nuclear magnetic resonance (NMR). MPC films are assembled on SAM modified gold electrode interfaces by using a "dip cycle" method of alternating MPC layers and dithiol linking molecules. Film growth at gold electrode is tracked electrochemically by measuring changes to the double layer charging current of the system. Analogous films assembled on silane modified glass slides allow for optical monitoring of film growth and cross-sectional TEM analysis provides an estimated film thickness. During film assembly, manipulation of the MPC ligand protection as well as the interparticle linkage mechanism allow for networked films, that are readily adaptable, to interface with redox protein having different adsorption mechanism. For example, Pseudomonas aeruginosa azurin (AZ) can be adsorbed hydrophobically to dithiol-linked films of hexanethiolate MPCs and cytochrome c (cyt c) can be immobilized electrostatically at a carboxylic acid modified MPC interfacial layer. In this report, we focus on the film protocol for the AZ system exclusively. Investigations involving the adsorption of proteins on MPC modified synthetic platforms could further the understanding of interactions between biomolecules and man-made materials, and consequently aid the development of biosensor schemes, ET modeling systems, and synthetic biocompatible materials.5-8

Alkanethiolate stabilized gold colloids known as monolayer protected clusters (MPCs) are synthesized, characterized, and assembled into thin films as an adsorption interface for protein monolayer electrochemistry of simple redox protein like Pseudomonas aeruginosa azurin (AZ) and cytochrome c (cyt c).

Monolayer의 합성, 조립, 그리고 특성화는 단백질 Monolayer의 전기에 대한 골드 Nanoparticle 필름을 보호

Colloidal gold nanoparticles protected with alkanethiolate ligands called monolayer protected gold clusters (MPCs) are synthesized and subsequently ...

Observing and Quantifying Fibroblast-mediated Fibrin Gel Compaction

Cells embedded in collagen and fibrin gels attach and exert traction forces on the fibers of the gel. These forces can lead to local and global reorganization and realignment of the gel microstructure. This process proceeds in a complex manner that is dependent in part on the interplay between the location of the cells, the geometry of the gel, and the mechanical constraints on the gel. To better understand how these variables produce global fiber alignment patterns, we use time-lapse differential interference contrast (DIC) microscopy coupled with an environmentally controlled bioreactor to observe the compaction process between geometrically spaced explants (clusters of fibroblasts). The images are then analyzed with a custom image processing algorithm to obtain maps of the strain. The information obtained from this technique can be used to probe the mechanobiology of various cell-matrix interactions, which has important implications for understanding processes in wound healing, disease development, and tissue engineering applications.

Time-lapse microscopy and image processing techniques were used to observe and analyze fibroblast-mediated gel compaction and fibrin fiber realignment in an environmentally controlled bioreactor over a 48 hr&#160;period.

섬유아세포 매개 피브린 겔 다짐 관찰 및 정량화

Cells embedded in collagen and fibrin gels attach and exert traction forces on the fibers of the gel. These forces can lead to local and global ...

Synthesis of an Intein-mediated Artificial Protein Hydrogel

We present the synthesis of a highly stable protein hydrogel mediated by a split-intein-catalyzed protein trans-splicing reaction. The building blocks of this hydrogel are two protein block-copolymers each containing a subunit of a trimeric protein that serves as a crosslinker and one half of a split intein. A highly hydrophilic random coil is inserted into one of the block-copolymers for water retention. Mixing of the two protein block copolymers triggers an intein trans-splicing reaction, yielding a polypeptide unit with crosslinkers at either end that rapidly self-assembles into a hydrogel. This hydrogel is very stable under both acidic and basic conditions, at temperatures up to 50 &#176;C, and in organic solvents. The hydrogel rapidly reforms after shear-induced rupture. Incorporation of a &#34;docking station peptide&#34; into the hydrogel building block enables convenient incorporation of &#34;docking protein&#34;-tagged target proteins. The hydrogel is compatible with tissue culture growth media, supports the diffusion of 20 kDa molecules, and enables the immobilization of bioactive globular proteins. The application of the intein-mediated protein hydrogel as an organic-solvent-compatible biocatalyst was demonstrated by encapsulating the horseradish peroxidase enzyme and corroborating its activity.

We present the synthesis of a split-intein-mediated protein hydrogel. The building blocks of this hydrogel are two protein copolymers each containing a subunit of a trimeric protein that serves as a crosslinker and one half of a split intein. Mixing of the two protein copolymers triggers an intein trans-splicing reaction, yielding a polypeptide unit that self-assembles into a hydrogel. This hydrogel is highly pH- and temperature-stable, compatible with organic solvents, and easily incorporates functional globular proteins.

테인 매개 인공 단백질 히드로 겔의 합성

We present the synthesis of a highly stable protein hydrogel mediated by a split-intein-catalyzed protein trans-splicing reaction. The building blocks of ...

Formation of Biomembrane Microarrays with a Squeegee-based Assembly Method

Lipid bilayer membranes form the plasma membranes of cells and define the boundaries of subcellular organelles. In nature, these membranes are heterogeneous mixtures of many types of lipids, contain membrane-bound proteins and are decorated with carbohydrates. In some experiments, it is desirable to decouple the biophysical or biochemical properties of the lipid bilayer from those of the natural membrane. Such cases call for the use of model systems such as giant vesicles, liposomes or supported lipid bilayers (SLBs). Arrays of SLBs are particularly attractive for sensing applications and mimicking cell-cell interactions. Here we describe a new method for forming SLB arrays. Submicron-diameter SiO2 beads are first coated with lipid bilayers to form spherical SLBs (SSLBs). The beads are then deposited into an array of micro-fabricated submicron-diameter microwells. The preparation technique uses a "squeegee" to clean the substrate surface, while leaving behind SSLBs that have settled into microwells. This method requires no chemical modification of the microwell substrate, nor any particular targeting ligands on the SSLB. Microwells are occupied by single beads because the well diameter is tuned to be just larger than the bead diameter. Typically, more 75% of the wells are occupied, while the rest remain empty. In buffer SSLB arrays display long-term stability of greater than one week. Multiple types of SSLBs can be placed in a single array by serial deposition, and the arrays can be used for sensing, which we demonstrate by characterizing the interaction of cholera toxin with ganglioside GM1. We also show that phospholipid vesicles without the bead supports and biomembranes from cellular sources can be arrayed with the same method and cell-specific membrane lipids can be identified.

Supported lipid bilayers and natural membrane particles are convenient systems that can approximate the properties of cell membranes and be incorporated in a variety of analytical strategies. Here we demonstrate a method for preparing microarrays composed of supported lipid bilayer-coated SiO2 beads, phospholipid vesicles or natural membrane particles.

스퀴지 기반의 조립 방법과 생체막 마이크로 어레이의 형성

Lipid bilayer membranes form the plasma membranes of cells and define the boundaries of subcellular organelles. In nature, these membranes are ...

Synthesis of Keratin-based Nanofiber for Biomedical Engineering

Electrospinning, due to its versatility and potential for applications in various fields, is being frequently used to fabricate nanofibers. Production of these porous nanofibers is of great interest due to their unique physiochemical properties. Here we elaborate on the fabrication of keratin containing poly (&#949;-caprolactone) (PCL) nanofibers (i.e., PCL/keratin composite fiber). Water soluble keratin was first extracted from human hair and mixed with PCL in different ratios. The blended solution of PCL/keratin was transformed into nanofibrous membranes using a laboratory designed electrospinning set up. Fiber morphology and mechanical properties of the obtained nanofiber were observed and measured using scanning electron microscopy and tensile tester. Furthermore, degradability and chemical properties of the nanofiber were studied by FTIR. SEM images showed uniform surface morphology for PCL/keratin fibers of different compositions. These PCL/keratin fibers also showed excellent mechanical properties such as Young&#39;s modulus and failure point. Fibroblast cells were able to attach and proliferate thus proving good cell viability. Based on the characteristics discussed above, we can strongly argue that the blended nanofibers of natural and synthetic polymers can represent an excellent development of composite materials that can be used for different biomedical applications.

Electrospun nanofibers have a high surface area to weight ratio, excellent mechanical integrity, and support cell growth and proliferation. These nanofibers have a wide range of biomedical applications. Here we fabricate keratin/ PCL nanofibers, using the electrospinning technique, and characterize the fibers for possible applications in tissue engineering.

의 생명 공학에 대한 각질 기반의 나노 섬유의 합성

Electrospinning, due to its versatility and potential for applications in various fields, is being frequently used to fabricate nanofibers. Production of ...

Synthesis of Gold Nanoparticle Integrated Photo-responsive Liposomes and Measurement of Their Microbubble Cavitation upon Pulse Laser Excitation

Photo-responsive nanoparticles (NPs) have received considerable attention because of their potential in providing spatial, temporal, and dosage control over the drug release. However, most of the relevant technologies are still in the development process and are unprocurable by clinics. Here, we describe a facile fabrication of these photo-responsive NPs with commercially available gold NPs and thermo-responsive liposomes. Calcein is used as a model drug to evaluate the encapsulation efficiency and the release kinetic profile upon heat/light stimulation. Finally, we show that this photo-triggered release is due to the membrane disruption caused by microbubble cavitation, which can be measured with hydrophone.

This protocol describes a simple preparation method for gold nanoparticle integrated photo-responsive liposomes with the commercially available materials. It also shows how to measure the microbubble cavitation process of the synthesized liposomes upon the treatment of pulsed laser.

펄스 레이저인가 전압에 따라 금 나노 입자 통합 사진 응답 리포좀과 그들의 마이크로 버블 공동 현상의 측정의 합성

Photo-responsive nanoparticles (NPs) have received considerable attention because of their potential in providing spatial, temporal, and dosage control ...

Synthesis of Functionalized 10-nm Polymer-coated Gold Particles for Endothelium Targeting and Drug Delivery

Gold nanoparticles (AuNPs) have been used extensively in medical research due to their size, biocompatibility, and modifiable surface. Specific targeting and drug delivery are some of the applications of these AuNPs, but endothelial extracellular matrices&#39; defensive properties hamper particle uptake. To address this issue, we describe a synthesis method for ultrasmall gold nanoparticles to improve vascular delivery, with customizable functional groups and polymer lengths for further adjustments. The protocol yields 2.5 nm AuNPs that are capped with tetrakis(hydroxymethyl)phosphonium chloride (THPC). The replacement of THPC with hetero-functional polyethylene glycol (PEG) on the surface of the AuNP increases the hydrodynamic radius to 10.5 nm while providing various functional groups on the surface. The last part of the protocol includes an optional addition of a fluorophore to allow the AuNPs to be visualized under fluorescence to track nanoparticle uptake. Dialysis and lyophilization were used to purify and isolate the AuNPs. These fluorescent nanoparticles can be visualized in both in vitro and in vivo experiments due to the biocompatible PEG coating and fluorescent probes. Additionally, the size range of these nanoparticles render them an ideal candidate for probing the glycocalyx without disrupting normal vasculature function, which may lead to improved delivery and therapeutics.

We describe a method of synthesizing biocompatible 10-nm gold nanoparticles, functionalized by coating poly-ethylene glycol onto the surface. These particles can be used in vitro and in vivo for delivering therapeutics to nanoscale cellular and extracellular spaces that are difficult to access with conventional nanoparticle sizes.

기능성된 10 nm 폴리머-코팅 된 금 입자 피 대상에 대 한 및 약물 전달의 합성

Gold nanoparticles (AuNPs) have been used extensively in medical research due to their size, biocompatibility, and modifiable surface. Specific targeting ...

Synthesis of Multi-walled Carbon Nanotubes Modified with Silver Nanoparticles and Evaluation of Their Antibacterial Activities and Cytotoxic Properties

In this study, multi-walled carbon nanotubes (MWCNTs) were treated with an aqueous sulfuric acid solution to form an oxygen-based functional group. Silver MWCNTs were prepared by the reductive deposition of silver from an aqueous solution of AgNO3 on the oxidized MWCNTs. Given the unique color of the CNTs, it was not possible to apply them to the minimum inhibitory concentration or mitochondrial toxicity assays to evaluate the toxicity and antibacterial properties, since they would interfere with the assays. The inhibition zone and minimum bactericidal concentration for the Ag-MWCNTs were measured and Live/Dead and Trypan Blue assays were used to measure the toxicity and antibacterial properties without interfering with the color of the CNTs.

In this study, antimicrobial nanomaterials were synthesized by acidic oxidation of multiwalled carbon nanotubes and subsequent reductive deposition of silver nanoparticles. Antimicrobial activity and cytotoxicity tests were performed with the as-prepared nanomaterials.

나노 입자와 그들의 항균 활동 및 세포 독성 속성의 평가 수정 다중 벽 탄소 나노튜브의 합성

In this study, multi-walled carbon nanotubes (MWCNTs) were treated with an aqueous sulfuric acid solution to form an oxygen-based functional group. Silver ...

In Vesiculo Synthesis of Peptide Membrane Precursors for Autonomous Vesicle Growth

Compartmentalization of biochemical reactions is a central aspect of synthetic cells. For this purpose, peptide-based reaction compartments serve as an attractive alternative to liposomes or fatty acid-based vesicles. Externally or within the vesicles, peptides can be easily expressed and simplify the synthesis of membrane precursors. Provided here is a protocol for the creation of vesicles with diameters of ~200 nm based on the amphiphilic elastin-like polypeptides (ELP) utilizing dehydration-rehydration from glass beads. Also presented are protocols for bacterial ELP expression and purification via inverse temperature cycling, as well as their covalent functionalization with fluorescent dyes. Furthermore, this report describes a protocol to enable the transcription of RNA aptamer dBroccoli inside ELP vesicles as a less complex example for a biochemical reaction. Finally, a protocol is provided, which allows in vesiculo expression of fluorescent proteins and the membrane peptide, whereas synthesis of the latter results in vesicle growth.

Presented here are protocols for the creation of peptide-based small unilamellar vesicles capable of growth. To facilitate in vesiculo production of the membrane peptide, these vesicles are equipped with a transcription-translation system and the peptide-encoding plasmid.

자율 적인 소포 성장을 위한 펩티드 멤브레인 전구체의 베시큘로 합성

Compartmentalization of biochemical reactions is a central aspect of synthetic cells. For this purpose, peptide-based reaction compartments serve as an ...