이 작품은 보조금의 원조 교육부, 문화, 스포츠, 일본의 과학 기술, 교토 Technoscience 센터 및 연구 개발에 대한 미타니 재단 젊은 과학자 (16K17956)에 의해 지원되었다.

1. 악기 <ol><li> 편광 장치 <ol><li> 편광 소자를 구성하는, 할로겐 램프, 도광판, 편광판, 시료 스테이지, 광학 레일로드 스탠드, 디지털 일안 리플렉스 카메라 (갖는 광원을 제공하는 편광에 대한도 1C와 자재 목록 참조 장치 부). </li></ol></li></ol> 생체 고분자의 용액의 제조 (2) <ol><li> 다당류 솔루션 <ol><li> 이상 ~ 12 시간 동안 80 ℃에서 교반하여 순수한 물 (100 mL) 중 sacran 7 (0.5 g)을 녹인다. 용해 중에, 증발을 방지하기 위해 플라스틱 포장으로 용기 커버. 동일하게 크 산탄 검의 수용액을 준비한다. </li><li> 0.5 중량 % 수용액을 얻었다 ~ 25 ℃에서 용액을 냉각. </li><li> 불순물 (48,400 XG, 4 ℃, 1 시간, 3 팀을 제거하는 용액을 원심 sacranES). </li></ol></li><li> MT 솔루션 <ol><li> 브리튼 - 로빈슨 완충액에서 0.5 중량 %의 튜 불린 용액 (1 ml)에 준비 (80 mm의 N, N &#39;비스은 피페 라진 (2- 에탄 설 폰산) (PIPES), 1 mM의 에틸렌 글리콜 비스 (β 아미노 에틸 에테르) - N, N, N &#39;, N&#39;-tetraacetic 아세트산 (EGTA); 얼음 8 5 밀리미터의 MgCl 2, pH를 6.8). </li><li> 0.5 중량 % 튜 불린 용액 (50 μL) 및 사용 구아노 신 - 5 &#39;- [(α, β) -methyleno] 포스페이트 (GpCpp) (5 μL)는 GpCpp 함유 튜 불린 용액 (50 μL)을 제조 하였다. 안정한 MT 핵을 얻기 위해 3 시간 동안 37 ℃에서 인큐베이션. 참고 : GpCpp의 역할은 MT 형성을 지원하기 위해 완전히 튜 불린에 MT의 해중합을 억제하는 것입니다. </li><li> 안정한 0.5 중량 % MT 용액을 수득 1 일 ~ 25 ° C에서 0.5 중량 % 튜 불린 용액 (950 μL) 및 GpCpp 튜 불린 - 함유 용액 (50 μL)를 섞는다. </li></ol></li><li> DNA 솔루션<ol><li> (1mM의 EDTA 10 개 mM 트리스, pH를 8.0) 트리스 EDTA 완충액에서 0.5 중량 %의 DNA 용액 (1 ml)로 준비한다. </li></ol></li><li> 건조 실험을 25 ℃에서 0.5 중량 %의 시료 생체 고분자 용액을 유지한다. </li></ol> 교차 편광 3. 건조 실험 및 관찰 <ol><li> 한 측면에서 개방 셀 솔루션 (도 1a) <ol><li> 실리콘 낱장 1mm의 두께를 적절한 형상으로 (자재 목록 참조) (5-15 mm의 1mm의 내부 크기 및 ~ 20mm;도 1a). <ol><li> ~ 20mm 두 비 개질 된 유리 슬라이드 (76mm × 1 mm × 26 mm)을 5-15 mm 1 mm의 내 치수를 갖는 실리콘으로 이루어지는 스페이서의 한쪽 개방 셀을 조립하고. 누출에서 시료 용액을 유지하기 위해 사전에 더블 클립 셀의 양면을 수정합니다. </li></ol></li><li> 천천히 ~ 1 mm의 기공 크기를 피펫을 사용하여 0.5 중량 %의 생체 고분자 용액 (100-300 μL)을 각각 추가~ 25 ℃에서 각 셀에 TE 팁. 주사기 바늘을 사용하여 세포로부터 기포를 제거한다. </li><li> 증발 대기압하에 60 ℃에서 공기 순환 오븐에서 함께 세포로; 증착 방향이 중력의 반대이다. </li></ol></li><li> 교차 편광하에 관찰 (도 1B-1C) <ol><li> 광역 (80mm X 80mm) 위에 평평한면 광원으로는 100 W 할로겐 램프를 통해 직선 가시 광선을 제공한다. 홀더 (도 1C)를 사용하여 45 ° 및 135 °에 편광판을 조정한다. </li><li> 광학 레일로드 스탠드 (도 1C)를 사용하여, 광원, 편광기, 샘플 스테이지, 카메라의 위치를 고정한다. 두 편광판 사이에 시료 스테이지를 배치 (편광판 사이의 거리는 5cm ~이어야한다). 집중 할 수 있도록 샘플 단계부터 ~ 20cm를 카메라를 놓습니다. </li><li> 주어진 시간에, 폴라 사이의 단계 3.1.3에서 샘플을 배치XZ 평면에 평행 한 단에 rizers 검은 커튼 장치를 커버; 실제 장치가도 1c에 도시되어있다. </li><li> 표준 줌 렌즈와 디지털 일안 리플렉스 카메라를 사용하여 선형 교차 편광판을 통해 샘플을 촬영 (재료 목록 참조). 컴퓨터 소프트웨어 (자료 목록을 참조)을 사용하여, 같은 초점 거리와 같은 카메라 설정을 제어 할 수 있습니다. </li></ol></li><li> 투과광 강도의 시공간적 분석 (도 1C) <ol><li> 건조 공정의 순서 배 양성 파라미터의 변화를 평가하기 위해 24 시간 동안 시간당 사진을 수집한다. </li><li> (예 ImageJ에) 화상 처리 프로그램을 사용하여 그레이 값과 Z 방향의 중심선을 따라 투과광의 강도를 측정한다. </li><li> 상부 개방 측으로부터의 거리의 함수로서 그레이 값의 그래프를 그린다. </li></ol></li><li> 교차 편광 조명 관리에서 현미경 관찰t (도 1D) <ol><li> , 방법을 확인 CCD 카메라 (9)가 장착 된 편광 현미경 현미경 관찰을합니다. 빛의 경로에 일차 위상차 판을 보관하십시오. PC에 소프트웨어를 사용하여 사진에 대한 조건을 제어합니다. </li></ol></li></ol>

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저자가 공개하는 게 없다.

이 때문에 투과광 강도가 너무 낮있는 카메라 샘플에 초점을 때로는 어려웠다. 이러한 경우, 무대에서 확장 된 투명 플라스틱 필름을 배치하는 것은 초점을 준비하는 데 도움이. 관측 가능한 해상도의 한계는 ~,이 경우에는 10 μm의 카메라 렌즈에 의존했다. 샘플의 두께, ΔY, 상기 관찰 ~ 제한이 경우 10mm 램프의 최대 광 강도에 의존했다. 도 1b에 도시 된 장치의 장?...

생체 고분자의 강성, 막대 형상의 구조체에 초점을 동적 부드러운 물질은 다당류 생물막 행렬 1, 세포 골격 단백질이 이루어지는 &quot;활성 겔&quot;을 원하는 형상 (3)의 &quot;DNA 접기」등, 다양한 용도로 사용되고있다. 구조적 특성을 명확히하기 위해, 많은 전략들이 이러한 투과형 전자 현미경, 주사 전자 현미경, 원자력 현미경, 및 공 초점 형광 현미경으로, 탐구되었다. 이러한 방법은 대부분 건조 또는 정적 상태에서 수행되기 때문에 그러나, 실제 생활 시스템에서 볼 수 있듯이, 거시적 규모의 동적 행동을 설명하기 어렵다. 최근에는 성공적으로 편광 된 광 (4)를 통해 수성 공기 LC 인터페이스 생체 고분자의 동적 거동을 관찰 하였다. 배향 구조의 시각화 동안 고분자 물질을 건조하는 동안용액의 시간적 변화는 불안정한 공기 LC 인터페이스 생체 고분자의자가 통합을 나타냈다. 여기서는 편광 수단을 이용하여, 공기 계면에서 LC LC 생체 고분자 용액의 건조를위한 프로토콜을 기술한다. 5,6 건조 고려하지 않는다는 LC상의 다른 분석 달리, 건조 공정 동안 LC 역학은 일측 개방 셀 내의 유체의 상 측면도의 배 양성 위해 파라미터를 평가하여 현재 조사했다 . 셀 증착의 조합 제어 증발 방향 육안 모니터링 허용 편광 수단의 사용. 또한,이 방법의 효과를 입증하기 위해, 분자량, 농도 등의 영향을받은 흡착 마이크로 도메인의 결정 구조에 집중하여 건조 기록을 확인할 수 있었다 건조 프로다당류, 미세 소관 (MTS), 및 DNA 등의 딱딱한 막대 모양 기본 생체 고분자의 세 스는 조사 하였다. 그들이 megamolecular 무게와 계층 거대 분자의 전형적인 예이며, 자신의 분자간 상호 작용 LC 상태를 형성 할 수 있도록하기 때문에 우리는이 생체 고분자를 선택했다.

<table><tbody><tr><td>sacran</td><td>Green Science Materials Inc., Japan</td><td></td><td>From Aphanothece sacrum.&lt;br/&gt; M&lt;sub&gt;w&lt;/sub&gt; = 1.9 &amp;times; 10&lt;sup&gt;7&lt;/sup&gt; g mol&lt;sup&gt;-1&lt;/sup&gt;</td></tr><tr><td>xanthan gum</td><td>Taiyo Kagaku Co., Japan</td><td>Neosoft XC</td><td>From Xanthomonas campestris.&lt;br/&gt; M&lt;sub&gt;w&lt;/sub&gt; = 4.7 &amp;times; 10&lt;sup&gt;6&lt;/sup&gt; g mol&lt;sup&gt;-1&lt;/sup&gt;</td></tr><tr><td>tubulin</td><td>Cytoskeleton, Inc., USA</td><td>T240</td><td>From porcine brain.</td></tr><tr><td>GpCpp</td><td>Jena Bioscience, Germany</td><td>NU405L</td><td></td></tr><tr><td>piperazine-&lt;em&gt;N,N&amp;prime;&lt;/em&gt;-bis(2-ethanesulfonic acid)</td><td>Sigma-Aldrichi, Co. LLC.</td><td>P6757-500G</td><td>PIPES</td></tr><tr><td>ethylene glycol-bis(&amp;beta;-aminoethyl ether)-&lt;em&gt;N,N,N&#039;,N&#039;&lt;/em&gt;-tetraacetic acid</td><td>Dojindo Molecular Technologies, Inc.</td><td>342-01314</td><td>EGTA</td></tr><tr><td>MgCl&lt;sub&gt;2&lt;/sub&gt;-6H&lt;sub&gt;2&lt;/sub&gt;O</td><td>Wako Pure Chemical Industries, Ltd.</td><td>135-15055</td><td></td></tr><tr><td>KOH</td><td>Wako Pure Chemical Industries, Ltd.</td><td>162-21813</td><td>pellet</td></tr><tr><td>DNA</td><td>Sigma-Aldrich, Co. LLC.</td><td>D1626</td><td>From salmon testes.&lt;br/&gt;Mw = 1.3 &amp;times; 10&lt;sup&gt;6&lt;/sup&gt; Da (~2,000 bp)</td></tr><tr><td>Tris-EDTA buffer solution</td><td>Sigma-Aldrich, Co. LLC.</td><td>T9285-100ML</td><td>10 mM Tris, pH 8.0, with 1 mM EDTA</td></tr><tr><td>slide glass</td><td>Matsunami Glass Ind., Ltd., Japan</td><td>S1111</td><td></td></tr><tr><td>silicon rubber sheet</td><td>Asone Co.</td><td>6-611-32</td><td>Thickness: 1 mm</td></tr><tr><td>centrifuge</td><td>Beckman-Coulter, Inc., USA</td><td>Avanti J-25</td><td>equipped with a JA-20 rotor</td></tr><tr><td>light source</td><td>Sumita Optical Glass, Inc., Japan</td><td>LS-LHA</td><td></td></tr><tr><td>light guide</td><td>Sumita Optical Glass, Inc., Japan</td><td>GF7.2-1-L1500R-M80 (AAAR-015M)</td><td>80 mm &amp;times; 80 mm</td></tr><tr><td>halogen lamp</td><td>Ushio Inc., Japan</td><td>JCR 15V150WBN</td><td></td></tr><tr><td>holder</td><td>Sigmakoki, Co.,Ltd.</td><td>KMH-80</td><td></td></tr><tr><td>sample stage</td><td>Sigmakoki, Co.,Ltd.</td><td>TARW-25503L</td><td></td></tr><tr><td>sample holder</td><td>Sigmakoki, Co.,Ltd.</td><td>SHA-25RO</td><td></td></tr><tr><td>rod</td><td>Sigmakoki, Co.,Ltd.</td><td>ROU-12-40</td><td></td></tr><tr><td>posts holder</td><td>Sigmakoki, Co.,Ltd.</td><td>RS-6-40</td><td></td></tr><tr><td>posts holder</td><td>Sigmakoki, Co.,Ltd.</td><td>RS-12-60</td><td></td></tr><tr><td>posts holder</td><td>Sigmakoki, Co.,Ltd.</td><td>RS-12-80</td><td></td></tr><tr><td>posts holder</td><td>Sigmakoki, Co.,Ltd.</td><td>RS-12-130</td><td></td></tr><tr><td>carrier</td><td>Sigmakoki, Co.,Ltd.</td><td>CAA-25LS</td><td></td></tr><tr><td>camera holder</td><td>Sigmakoki, Co.,Ltd.</td><td>CMH-2</td><td></td></tr><tr><td>medium optical rail</td><td>Sigmakoki, Co.,Ltd.</td><td>OBA-500SH</td><td></td></tr><tr><td>lenstube</td><td>Tomytech, BORG</td><td>lenstube BK</td><td>80&amp;phi;, L25 mm&amp;nbsp;</td></tr><tr><td>lenstube</td><td>Tomytech, BORG</td><td>lenstube BK</td><td>80&amp;phi;, L50 mm&amp;nbsp;</td></tr><tr><td>multiband</td><td>Tomytech, BORG</td><td></td><td>80&amp;phi;&amp;nbsp;</td></tr><tr><td>V plate</td><td>Tomytech, BORG</td><td>V plate 60S</td><td></td></tr><tr><td>plate holder</td><td>Viexen, Co.,Ltd.</td><td>plate holder SX</td><td></td></tr><tr><td>EOS Kiss X7i&amp;nbsp;</td><td>Canon Inc., Japan</td><td>8594B001</td><td>with a standard zoom lens,&amp;nbsp; EFP 18-55 mm</td></tr><tr><td>photographic software</td><td>Canon Inc., Japan</td><td>EOS Utility</td><td></td></tr><tr><td>PC</td><td>Microsoft</td><td>Surface</td><td></td></tr><tr><td>polarization microscope</td><td>Olympus</td><td>BX51</td><td></td></tr><tr><td>first order retardation plate</td><td>Olympus</td><td>U-TP530</td><td>&amp;lambda; = 530 nm</td></tr><tr><td>CCD camera</td><td>Olympus</td><td>DP80</td><td></td></tr><tr><td>photographic software</td><td>Olympus</td><td>cellSens Standard</td><td></td></tr><tr><td>Java-based image processing program</td><td>the National Institutes of Health</td><td>ImageJ</td><td></td></tr></tbody></table>

methods for the self-integration of megamolecular biopolymers on the drying air-lc interface

물을 사용 살아있는 유기체는 항상 환경에서 건조하는 경향이 있습니다. 관다발 물을 이동 및 피부 층에 물 보습의 경우에서 볼 수 있듯이 이들의 활동은 생체 고분자 기반의 마이크로 및 매크로 구조에 의해 구동된다. 본 연구에서는 건조 생체 고분자로 이루어지는 수계 결정 (LC) 용액의 효과를 평가하기위한 방법을 개발 하였다. LC의 바이오 폴리머가 megamolecular 무게를 가지고, 우리는 다당류, 세포 골격 단백질과 DNA를 연구하기로 결정했습니다. 편광하에 건조시 생체 고분자 용액의 관찰 불안정한 공기 LC 인터페이스부터 milliscale자가 통합을 보여준다. 수성 LC 생체 고분자 용액의 역학은 일측 개방 셀로부터 물을 증발에 의해 모니터링 될 수있다. 교차 편광을 이용하여 촬영 된 영상을 분석함으로써, 배 양성 위해 파라미터의 시공간적 변화를 인식 할 수있다. 이방법은 인공 다양한 분야에서 물질뿐만 아니라 자연의 살아있는 조직뿐만 아니라의 특성 유용 할 수 있습니다. 우리는 생물 의학 및 환경 분야에서의 부드러운 소재에 대한 평가 방법을 제공 할 것입니다 믿습니다.

<html> 도 1에 도시 된 바와 같은 장치를 사용하여 미소 영역에서 자기 통합 에어 LC 인터페이스 평가 건조 (도 2A)에 macrodomain한다. 건조 실험의 처음 입증 된 바와 같이, megamolecular 다당류 sacran (M w = 1.9 × 10 7g 몰 -1) 및 크 산탄 검 (4.7 × 106 그램 몰 -1) 두 종류, 비교 하였다. 도 2b는 교차 편광?...

Watch this Scientific Journal Video about Methods for the Self-integration of Megamolecular Biopolymers on the Drying Air-LC Interface at JoVE.com

Methods for the Self-integration of Megamolecular Biopolymers on the Drying Air-LC Interface

공기 액정 성 인터페이스 megamolecular 생체 고분자의 건조 - 유도 된자가 통합을위한 방법이 여기에 제공된다. 이 방법은 생체 고분자의 거시적 인 잠재력을 이해뿐만 아니라, 생물 의학, 환경 분야의 부드러운 소재에 대한 평가 방법으로뿐만 아니라 도움이 될 것입니다.

건조 공기-LC 인터페이스에 Megamolecular 생체 고분자의 자체 통합을위한 방법

Living organisms that use water are always prone to drying in the environment. Their activities are driven by biopolymer-based micro- and ...

methods-for-self-integration-megamolecular-biopolymers-on-drying-air

Research

JoVE Journal

Bioengineering

6.0K 조회수.  Japan Advanced Institute of Science and Technology.  공기 액정 성 인터페이스 megamolecular 생체 고분자의 건조 - 유도 된자가 통합을위한 방법이 여기에 제공된다. 이 방법은 생체 고분자의 거시적 인 잠재력을 이해뿐만 아니라, 생물 의학, 환경 분야의 부드러운 소재에 대한 평가 방법으로뿐만 아니라 도움이 될 것입니다. 

비디오: Methods for the Self-integration of Megamolecular Biopolymers on the Drying Air-LC Interface

Synthesis of Poly(N-isopropylacrylamide) Janus Microhydrogels for Anisotropic Thermo-responsiveness and Organophilic/Hydrophilic Loading Capability

Janus microparticles are compartmentalized particles with differing molecular structures and/or functionality on each of their two sides. Because of this unique property, Janus microparticles have been recognized as a new class of materials, thereby attracting a great deal of attention from various research fields. The versatility of these microparticles has been exemplified through their uses as building blocks for self-assembly, electrically responsive actuators, emulsifiers for painting and cosmetics, and carriers for drug delivery. This study introduces a detailed protocol that explicitly describes a synthetic method for designing novel Janus microhydrogels composed of a single base material, poly(N-isopropylacrylamide) (PNIPAAm). Janus microdroplets are firstly generated via a hydrodynamic focusing microfluidic device (HFMD) based on the separation of a supersaturated aqueous NIPAAm monomer solution and subsequently polymerized through exposure to UV irradiation. The resulting Janus microhydrogels were found to be entirely composed of the same base material, featured an easily identifiable compartmentalized morphology, and exhibited anisotropic thermo-responsiveness and organophilic/hydrophilic loading capability. We believe that the proposed method introduces a novel hydrogel platform with the potential for advanced synthesis of multi-functional Janus microhydrogels.

Synthesis of PolyN-isopropylacrylamide Janus Microhydrogels for Anisotropic Thermo-responsiveness and Organophilic/Hydrophilic Loading Capability

We present a protocol to synthesize Janus microhydrogels composed entirely of the same base material, poly(N-isopropylacrylamide) (PNIPAAm), with a clearly compartmentalized structure base on the phase separation of a supersaturated NIPAAm monomer solution. The synthesized Janus microhydrogels show unique properties such as anisotropic thermo-responsiveness and organophilic/hydrophilic loading capability.

폴리의 합성 (<em&gt; N</em&gt; 이소 프로필 아크릴 아미드) 이방성 열 응답 성 및 유기화 / 친수성로드 기능에 대한 야누스 Microhydrogels

Janus microparticles are compartmentalized particles with differing molecular structures and/or functionality on each of their two sides. Because of this ...

연구

화학

Orientational Transition in a Liquid Crystal Triggered by the Thermodynamic Growth of Interfacial Wetting Sheets

In liquid crystal (LC) physical chemistry, molecules near the surface play a great role in controlling bulk orientation. Thus far, mainly to achieve desired molecular orientation states in LC displays, the "static" surface property of LCs, so-called surface anchoring, has been intensively studied. As a rule of thumb, once the initial orientation of LCs is "locked" by specific surface treatments, such as rubbing or treatment with a specific alignment layer, it hardly changes with temperature. Here, we present a system exhibiting an orientational transition upon temperature variation, which conflicts with the consensus. Right on the transition, the bulk LC molecules experience the orientational rotation, with 90&#176; between the planar (P) orientation at high temperatures and the vertical (V) orientation at low temperatures in the first-order transitional manner. We have tracked thermodynamic surface anchoring behavior by means of polarizing optical microscopy (POM), dielectric spectroscopy (DS), high-resolution differential scanning calorimetry (HR-DSC), and grazing incidence X-ray diffraction (GI-XRD) and reached a plausible physical explanation: that the transition is triggered by a growth of surface wetting sheets, which impose the V orientation locally against the P orientation in the bulk. This landscape would provide a general link explaining how the equilibrium bulk orientation is affected by surface-localized orientation in many LC systems. In our characterization, POM and DS are advantageous by offering information on the spatial distribution of the orientation of LC molecules. HR-DSC gives information about the precise thermodynamic information on transitions, which cannot be addressed by conventional DSC instruments due to limited resolution. GI-XRD provides information on surface-specific molecular orientation and short-range orderings. The goal of this paper is to present a protocol for preparing a sample that exhibits the transition and to demonstrate how the thermodynamic structural variation, both in the bulk and on surfaces, can be analyzed through the abovementioned methods.

Here, we present a protocol to trigger an orientational transition of a liquid crystal in response to temperature. Methodologies are described for preparing a sample in order to observe the transition and the detailed transitional evolution.

계면 습윤지의 열역학적 성장에 의해 촉발 된 액정의 방향 전환

In liquid crystal (LC) physical chemistry, molecules near the surface play a great role in controlling bulk orientation. Thus far, mainly to achieve ...

공학

Interfacial Molecular-level Structures of Polymers and Biomacromolecules Revealed via Sum Frequency Generation Vibrational Spectroscopy

As a second-order nonlinear optical spectroscopy, sum frequency generation (SFG) vibrational spectroscopy has widely been used in investigating various surfaces and interfaces. This non-invasive optical technique can provide the local molecular-level information with monolayer or submonolayer sensitivity. We here are providing experimental methodology on how to selectively detect the buried interface for both macromolecules and biomacromolecules. With this in mind, interfacial secondary structures of silk fibroin and water structures around model short-chain oligonucleotide duplex are discussed. The former shows a chain-chain overlap or spatial confinement effect and the latter shows a protection function against the Ca2+ ions resulting from the chiral spine superstructure of water.

Being comprehensively utilized, sum frequency generation (SFG) vibrational spectroscopy can help to reveal chain conformational order and secondary structural change happening at polymer and biomacromolecule interfaces.

합계 주파수 생성 진동 분광법을 통해 밝혀진 중합체 및 생체 거력의 계면 분자 수준 구조

As a second-order nonlinear optical spectroscopy, sum frequency generation (SFG) vibrational spectroscopy has widely been used in investigating various ...

생화학

Preparation of Liquid Crystal Networks for Macroscopic Oscillatory Motion Induced by Light

A strategy based on doped liquid crystalline networks is described to create mechanical self-sustained oscillations of plastic films under continuous light irradiation. The photo-excitation of dopants that can quickly dissipate light into heat, coupled with anisotropic thermal expansion and self-shadowing of the film, gives rise to the self-sustained deformation. The oscillations observed are influenced by the dimensions and the modulus of the film, and by the directionality and intensity of the light. The system developed offers applications in energy conversion and harvesting for soft-robotics and automated systems.
The general method described here consists of creating free-standing liquid crystalline films and characterizing the mechanical and thermal effects observed. The molecular alignment is achieved using alignment layers (rubbed polyimide), commonly used in the display manufacturing industry. To obtain actuators with large deformation, the mesogens are aligned and polymerized in a splay/bend configuration, i.e., with the director of the liquid crystals (LCs) going gradually from planar to homeotropic through the film thickness. Upon irradiation, the mechanical and thermal oscillations obtained are monitored with a high-speed camera. The results are further quantified by image analysis using an image processing program.

The goal of the protocol is to create liquid crystalline polymer films that can mechanically oscillate under continuous light irradiation. We describe in great detail the conception of free-standing films, from the liquid crystal alignment method to the photo-actuation. The experimental protocol applied to prepare this material is broadly applicable.

빛에 의해 유도 된 거시적인 진동 모션 액정 네트워크의 준비

A strategy based on doped liquid crystalline networks is described to create mechanical self-sustained oscillations of plastic films under continuous ...

Microfluidic Preparation of Liquid Crystalline Elastomer Actuators

This paper focuses on the microfluidic process (and its parameters) to prepare actuating particles from liquid crystalline elastomers. The preparation usually consists in the formation of droplets containing low molar mass liquid crystals at elevated temperatures. Subsequently, these particle precursors are oriented in the flow field of the capillary and solidified by a crosslinking polymerization, which produces the final actuating particles. The optimization of the process is necessary to obtain the actuating particles and the proper variation of the process parameters (temperature and flow rate) and allows variations of size and shape (from oblate to strongly prolate morphologies) as well as the magnitude of actuation. In addition, it is possible to vary the type of actuation from elongation to contraction depending on the director profile induced to the droplets during the flow in the capillary, which again depends on the microfluidic process and its parameters. Furthermore, particles of more complex shapes, like core-shell structures or Janus particles, can be prepared by adjusting the setup. By the variation of the chemical structure and the mode of crosslinking (solidification) of the liquid crystalline elastomer, it is also possible to prepare actuating particles triggered by heat or UV-vis irradiation.

This article describes the microfluidic process and parameters to prepare actuating particles from liquid crystalline elastomers. This process allows the preparation of actuating particles and the variation of their size and shape (from oblate to strongly prolate, core-shell, and Janus morphologies) as well as the magnitude of actuation.

액체 결정 탄성 액츄에이터의 미세 준비

This paper focuses on the microfluidic process (and its parameters) to prepare actuating particles from liquid crystalline elastomers. The preparation ...

Automated Lipid Bilayer Membrane Formation Using a Polydimethylsiloxane Thin Film

An artificial lipid bilayer, or black lipid membrane (BLM), is a powerful tool for studying ion channels and protein interactions, as well as for biosensor applications. However, conventional BLM formation techniques have several drawbacks and they often require specific expertise and laborious processes. In particular, conventional BLMs suffer from low formation success rates and inconsistent membrane formation time. Here, we demonstrate a storable and transportable BLM formation system with controlled thinning-out time and enhanced BLM formation rate by replacing conventionally used films (polytetrafluoroethylene, polyoxymethylene, polystyrene) to polydimethylsiloxane (PDMS). In this experiment, a porous-structured polymer such as PDMS thin film is used. In addition, as opposed to conventionally used solvents with low viscosity, the use of squalene permitted a controlled thinning-out time via slow solvent absorption by PDMS, prolonging membrane lifetime. In addition, by using a mixture of squalene and hexadecane, the freezing point of the lipid solution was increased (~16 &#176;C), in addition, membrane precursors were produced that can be indefinitely stored and readily transported. These membrane precursors have reduced BLM formation time of &#60; 1 hr and achieved a BLM formation rate of ~80%. Moreover, ion channel experiments with gramicidin A demonstrated the feasibility of the membrane system.

We demonstrate a storable, transportable lipid bilayer formation system. A lipid bilayer membrane can be formed within 1 hr with over 80% success rate when a frozen membrane precursor is brought to ambient temperature. This system will reduce laborious processes and expertise associated with ion channels.

폴리 디메틸 실록산 박막을 사용하여 자동화 지질 이중층 제막

An artificial lipid bilayer, or black lipid membrane (BLM), is a powerful tool for studying ion channels and protein interactions, as well as for ...

생체공학

Self-Assembly of Hybrid Lipid Membranes Doped with Hydrophobic Organic Molecules at the Water/Air Interface

Because of their unique properties, including an ultrathin thickness (3-4 nm), ultrahigh resistivity, fluidity and self-assembly ability, lipid bilayers can be readily functionalized and have been used in various applications such as bio-sensors and bio-devices. In this study, we introduced a planar organic molecule: copper (II) 2,9,16,23-tetra-tert-butyl-29H,31H-phthalocyanine (CuPc) to dope lipid membranes. The CuPc/lipid hybrid membrane forms at the water/air interface by self-assembly. In this membrane, the hydrophobic CuPc molecules are located between the hydrophobic tails of lipid molecules, forming a lipid/CuPc/lipid sandwich structure. Interestingly, an air-stable hybrid lipid bilayer can be readily formed by transferring the hybrid membrane onto a Si substrate. We report a straightforward method for incorporating nanomaterials into a lipid bilayer system, which represents a new methodology for the fabrication of biosensors and biodevices.

We report a protocol for producing a hybrid lipid membrane at the water/air interface by doping the lipid bilayer with copper (II) 2,9,16,23-tetra-tert-butyl-29H,31H-phthalocyanine (CuPc) molecules. The resulting hybrid lipid membrane has a lipid/CuPc/lipid sandwich structure. This protocol can also be applied to the formation of other functional nanomaterials.

물/공기 인터페이스에서 소수성 유기 분자와 함께 도핑 하이브리드 지질 막의 자체 조립

Because of their unique properties, including an ultrathin thickness (3-4 nm), ultrahigh resistivity, fluidity and self-assembly ability, lipid bilayers ...

Crystallization of Proteins on Chip by Microdialysis for In Situ X-ray Diffraction Studies

This protocol describes the manufacturing of reproducible and inexpensive microfluidic devices covering the whole pipeline for crystallizing proteins on-chip with the dialysis method and allowing in situ single-crystal or serial crystallography experiments at room temperature. The protocol details the fabrication process of the microchips, the manipulation of the on-chip crystallization experiments and the treatment of the in situ collected X-ray diffraction data for the structural elucidation of the protein sample. The main feature of this microfabrication procedure lies on the integration of a commercially available, semipermeable regenerated cellulose dialysis membrane in between two layers of the chip. The molecular weight cut-off of the embedded membrane varies depending on the molecular weight of the macromolecule and the precipitants. The device exploits the advantages of microfluidic technology, such as the use of minute volumes of samples (&#60;1 &#181;L) and fine tuning over transport phenomena. The chip coupled them with the dialysis method, providing precise and reversible control over the crystallization process and can be used for investigating phase diagrams of proteins at the microliter scale. The device is patterned using a photocurable thiolene-based resin with soft imprint lithography on an optically transparent polymeric substrate. Moreover, the background scattering of the materials composing the microchips and generating background noise was evaluated rendering the chip compatible for in situ X-ray diffraction experiments. Once protein crystals are grown on-chip up to an adequate size and population uniformity, the microchips can be directly mounted in front of the X-ray beam with the aid of a 3D printed holder. This approach addresses the challenges rising from the use of cryoprotectants and manual harvesting in conventional protein crystallography experiments through an easy and inexpensive manner. Complete X-ray diffraction data sets from multiple, isomorphous lysozyme crystals grown on-chip were collected at room temperature for structure determination.

Crystallization of Proteins on Chip by Microdialysis for In Situ X-ray Diffraction Studies

This paper details the fabrication protocol of microfluidic chips developed for on-chip protein crystallization with the dialysis method and in situ X-ray diffraction experiments. The microfabrication process makes it possible to integrate a semipermeable regenerated cellulose dialysis membrane with any molecular weight cut-off, between two layers of the chip.

시투 X선 회절 연구를 위한 미세 투약에 의한 칩에 단백질의 결정화

This protocol describes the manufacturing of reproducible and inexpensive microfluidic devices covering the whole pipeline for crystallizing proteins ...

Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes

Cellular membranes are highly crowded environments for biomolecular reactions and signaling. Yet, most in vitro experiments probing protein interaction with lipids employ naked bilayer membranes. Such systems lack the complexities of crowding by membrane-embedded proteins and glycans and exclude the associated volume effects encountered on cellular membrane surfaces. Also, the negatively charged glass surface onto which the lipid bilayers are formed prevents the free diffusion of transmembrane biomolecules. Here, we present a well-characterized polymer-lipid membrane as a mimic for crowded lipid membranes. This protocol utilizes polyethylene glycol (PEG)-conjugated lipids as a generalized approach for incorporating crowders into the supported lipid bilayer&#160;(SLB). First, a cleaning procedure of the microscopic slides and coverslips for performing single-molecule experiments is presented. Next, methods for characterizing the PEG-SLBs and performing single-molecule experiments of the binding, diffusion, and assembly of biomolecules using single-molecule tracking and photobleaching are discussed. Finally, this protocol demonstrates how to monitor the nanopore assembly of bacterial pore-forming toxin Cytolysin A (ClyA) on crowded lipid membranes with single-molecule photobleaching analysis. MATLAB codes with example datasets are also included to perform some of the common analyses such as particle tracking, extracting diffusive behavior, and subunit counting.

Here, a protocol to perform and analyze the binding, mobility, and assembly of single molecules on artificial crowded lipid membranes using single-molecule total internal reflection fluorescence (smTIRF) microscopy is presented.

단일 분자 확산 및 폴리머가 밀집된 지질 막에서의 조립

Cellular membranes are highly crowded environments for biomolecular reactions and signaling. Yet, most in vitro experiments probing protein interaction ...

Crystallization of Membrane Proteins in Lipidic Mesophases

Membrane proteins perform critical functions in living cells related to signal transduction, transport and energy transformations, and, as such, are implicated in a multitude of malfunctions and diseases. However, a structural and functional understanding of membrane proteins is strongly lagging behind that of their soluble partners, mainly, due to difficulties associated with their solubilization and generation of diffraction quality crystals. Crystallization in lipidic mesophases (also known as in meso or LCP crystallization) is a promising technique which was successfully applied to obtain high resolution structures of microbial rhodopsins, photosynthetic proteins, outer membrane beta barrels and G protein-coupled receptors. In meso crystallization takes advantage of a native-like membrane environment and typically produces crystals with lower solvent content and better ordering as compared to traditional crystallization from detergent solutions. The method is not difficult, but requires an understanding of lipid phase behavior and practice in handling viscous mesophase materials. Here we demonstrate a simple and efficient way of making LCP and reconstituting a membrane protein in the lipid bilayer of LCP using a syringe mixer, followed by dispensing nanoliter portions of LCP into an assay or crystallization plate, conducting pre-crystallization assays and harvesting crystals from the LCP matrix. These protocols provide a basic guide for approaching in meso crystallization trials; however, as with any crystallization experiment, extensive screening and optimization are required, and a successful outcome is not necessarily guaranteed.

The protocols describe the essential steps for obtaining diffraction quality crystals of a membrane protein starting from reconstitution of the protein in a lipidic cubic phase (LCP), finding initial conditions with LCP-FRAP pre-crystallization assays, setting up LCP crystallization trials and harvesting crystals.

Lipidic Mesophases의 막 단백질의 결정화

Membrane proteins perform critical functions in living cells related to signal transduction, transport and energy transformations, and, as such, are ...