우리는 없습니다 승인.

주의: 연구실 코트, 장갑, 그리고 안전 유리를 착용 하 고 사용 하기 전에 물질 안전 데이터 시트 (MSDS) 읽고 하십시오. 모든 구입한 재료 더 정화 없이 사용 되었습니다.
1. 스 티 렌/PFO/수성 SDS 솔루션 혼합물의 구성 된 유화 액의 준비
<ol><li>SDS 수 용액의 5mm를 준비 하려면 분해 순도 H2o.의 10 mL에 SDS의 14.5 밀리 그램 참고: 고 순도 H2O 사용 되었습니다 (저항 (R) = 18 m ω c m, 표면 장력 (γ) = 72.0 mN m-1 25 ° C에서).</li>
	<li>스 티 렌의 1.5 g, PFO, 0.6 g 및 SDS 수 용액의 5mm의 0.9 g 볶음 바 10ml 유리 약 병을 추가 합니다. 참고: 공초점 형광 현미경 관찰, 검색에 추가 2.6 coumarin 102의와 0.062 mg calcein의 혼합물에.</li>
	<li>실 온에서 1150 rpm에서 60 분을 위한 혼합물을 저 어 하 고 그 후 80 ° c 온도 높이</li>
	<li>1150 rpm에서 60 분 및 80 ° c.에 대 한 혼합물을 저 어</li>
</ol>2. 유화 액으로 구성 된의 스 티 렌/PFO/수성 SDS 솔루션 혼합물을 사용 하 여 고분자 입자의 제조
<ol><li>저 어 바 10ml 유리 약 병에 칼륨 peroxodisulfate의 3.9 mg, pyrene의 2 밀리 그램, 스 티 렌의 1.5 g, PFO, 0.6 g 및 SDS 수 용액의 5mm의 0.9 g을 추가 하 고 함께 고무 격 막 인감.</li>
	<li>30 분 동안 주사기 바늘을 통해 버블링 질소 가스 혼합물을 deoxygenate. 주의: 천천히 많은 양의 거품을 생성 하지에 질소 가스 흐름.</li>
	<li>준비 하는 유제, 1150 rpm 및 실 온에서 60 분을 위한 혼합물을 저 어 고 다음 80 ° c 온도 높이</li>
	<li>1150 rpm에서 30 분, 80 ° c.에 대 한 혼합물을 저 어</li>
	<li>테스트 튜브에 결과 솔루션 (3 mL)에 흐린 부분 (1.8 mL)를 전송 및 다음 추가 30% 수성 에탄올 솔루션 완전히 중 합 반응 종료.</li>
	<li>Sonicate (파워: 130 W, 주파수: 4.2 kHz) 결과 폴리머 입자를 세척 하 고 2300 x g에서 10 분 원심 다음 10 분.</li>
	<li>고분자 입자를, 테스트 튜브에서 표면에 뜨는 솔루션을 제거 합니다.</li>
	<li>테스트 튜브에 결과 고체를 물 3 mL를 추가 합니다. Sonicate (파워: 130 W, 주파수: 4.2 kHz) 10 분 및 2300 x g.에서 10 분 원심 분리기 테스트 튜브에서 표면에 뜨는 솔루션을 제거 하는 동안.</li>
	<li>(단계 2.8) 세척 절차를 반복 하 여 거품 표면에 뜨는 솔루션에서 생성 되지 않습니다. 투명 폴리머 입자의 고체를 물 증발.</li>
</ol>3. 투명 폴리머 입자를 사용 하 여 microcapsules의 준비
<ol><li>빈 폴리스 티 렌 입자 (단계 2.9)의 1 밀리 그램을 추가 저 어 바 10ml 유리 약 병에 물을 4 ml.</li>
	<li>Sonicate (파워: 130 W, 주파수: 4.2 kHz) 10 분 물에 빈 입자 분산.</li>
	<li>0.1 mL 물에 톨루엔의 분산 입자를 추가 하 고 실 온에서 100 rpm에서 1 h 동안 그것을 저 어. 테스트 튜브에 액체를 전송. 주의: 분당 회전 증가, 입자의 형태 변형 됩니다.</li>
	<li>2300 x에 10 분 동안 액체 원심 분리는 microcapsules에 그것을 형성. 테스트 튜브에서 표면에 뜨는 솔루션을 제거 합니다.</li>
</ol>

<ol><li>Okubo, M., Fujibayashi, T., Terada, A. <a target="_blank" href="https://link-springer-com.remotexs.ntu.edu.sg/article/10.1007/s00396-004-1210-4">Synthesis of micron-sized, monodisperse polymer particles of disc-like and polyhedral shapes by seeded dispersion polymerization.</a> Colloid Polym Sci. 283 (7), 793-798 (2005).</li>
<li>Fujibayashi, T., Okubo, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Preparation+and+Thermodynamic+Stability+of+Micron-Sized%2C+Monodisperse+Composite+Polymer+Particles+of+Disc-like+Shapes+by+Seeded+Dispersion+Polymerization%5BTitle%5D)+AND+%22Langmuir%22%5BJournal%5D)">Preparation and Thermodynamic Stability of Micron-Sized, Monodisperse Composite Polymer Particles of Disc-like Shapes by Seeded Dispersion Polymerization.</a> Langmuir. 23 (15), 7958-7962 (2007).</li>
<li>Okubo, M., Fujibayashi, T., Yamada, M., Minami, H. <a target="_blank" href="https://link-springer-com.remotexs.ntu.edu.sg/article/10.1007/s00396-004-1240-y">Micron-sized, monodisperse, snowman/confetti-shaped polymer particles by seeded dispersion polymerization.</a> Colloid Polym Sci. 283 (9), 1041-1045 (2005).</li>
<li>Xu, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Generation+of+Monodisperse+Particles+by+Using+Microfluidics%3A+Control+over+Size%2C+Shape%2C+and+Composition%5BTitle%5D)+AND+%22Angew+Chem+Int+Ed%22%5BJournal%5D)">Generation of Monodisperse Particles by Using Microfluidics: Control over Size, Shape, and Composition.</a> Angew Chem Int Ed. 44 (5), 724-728 (2005).</li>
<li>Serra, C. A., Chang, Z. <a target="_blank" href="http://onlinelibrary.wiley.com/doi/10.1002/ceat.200800219/abstract">Microfluidic-Assisted Synthesis of Polymer Particles.</a> Chem Eng Tech. 31 (8), 1099-1115 (2008).</li>
<li>Higuchi, T., Yabu, H., Shimomura, M. <a target="_blank" href="https://www-sciencedirect-com.remotexs.ntu.edu.sg/science/article/pii/S0927775705008101">Simple preparation of hemispherical polystyrene particles.</a> Colloids Surf A. 284, 250-253 (2006).</li>
<li>Sun, Z. Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Nonspherical+Colloidal+Crystals+Fabricated+by+the+Thermal+Pressing+of+Colloidal+Crystal+Chips%5BTitle%5D)+AND+%22Langmuir%22%5BJournal%5D)">Nonspherical Colloidal Crystals Fabricated by the Thermal Pressing of Colloidal Crystal Chips.</a> Langmuir. 21 (20), 8987-8991 (2005).</li>
<li>Deng, Y., et al. <a target="_blank" href="http://pubs.rsc.org/en/content/articlelanding/2008/jm/b714288a">A novel approach to the construction of 3-D ordered macrostructures with polyhedral particles.</a> J Mater Chem. 18 (4), 408-415 (2008).</li>
<li>Hyuk Im, S., Jeong, U., Xia, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Polymer+hollow+particles+with+controllable+holes+in+their+surfaces%5BTitle%5D)+AND+%22Nat+Mater%22%5BJournal%5D)">Polymer hollow particles with controllable holes in their surfaces.</a> Nat Mater. 4 (9), 671-675 (2005).</li>
<li>Saito, N., Kagari, Y., Okubo, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effect+of+Colloidal+Stabilizer+on+the+Shape+of+Polystyrene%2FPoly%28methyl+methacrylate%29+Composite+Particles+Prepared+in+Aqueous+Medium+by+the+Solvent+Evaporation+Method%5BTitle%5D)+AND+%22Langmuir%22%5BJournal%5D)">Effect of Colloidal Stabilizer on the Shape of Polystyrene/Poly(methyl methacrylate) Composite Particles Prepared in Aqueous Medium by the Solvent Evaporation Method.</a> Langmuir. 22 (22), 9397-9402 (2006).</li>
<li>Cai, P. -j, Tang, Y. -j, Wang, Y. -t, Cao, Y. -j <a target="_blank" href="https://www-sciencedirect-com.remotexs.ntu.edu.sg/science/article/pii/S0254058410004694">Fabrication of polystyrene hollow spheres in W/O/W multiple emulsions.</a> Mater Chem Phys. 124 (1), 10-12 (2010).</li>
<li>Liang, S. -S., Chen, S. -L., Chen, S. -H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Diverse+macroporous+spheres+synthesized+by+multiple+emulsion+polymerization+for+protein+analyses%5BTitle%5D)+AND+%22Chem+Commun%22%5BJournal%5D)">Diverse macroporous spheres synthesized by multiple emulsion polymerization for protein analyses.</a> Chem Commun. 47 (29), 8385-8387 (2011).</li>
<li>Kondo, Y., Yoshino, N. <a target="_blank" href="https://www-sciencedirect-com.remotexs.ntu.edu.sg/science/article/pii/S1359029405000312">Hybrid fluorocarbon/hydrocarbon surfactants.</a> Curr Opin Colloid Interface Sci. 10 (3-4), 88-93 (2005).</li>
<li>Takahashi, Y., Kondo, Y., Schmidt, J., Talmon, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Self-Assembly+of+a+Fluorocarbon-Hydrocarbon+Hybrid+Surfactant%3A+Dependence+of+Morphology+on+Surfactant+Concentration+and+Time%5BTitle%5D)+AND+%22J+Phys+Chem+B%22%5BJournal%5D)">Self-Assembly of a Fluorocarbon-Hydrocarbon Hybrid Surfactant: Dependence of Morphology on Surfactant Concentration and Time.</a> J Phys Chem B. 114 (42), 13319-13325 (2010).</li>
<li>Takahashi, Y., Nasu, Y., Aramaki, K., Kondo, Y. <a target="_blank" href="https://www-sciencedirect-com.remotexs.ntu.edu.sg/science/article/pii/S0022113912003429">Unusual viscoelastic behavior of aqueous solutions of fluorocarbon-hydrocarbon hybrid surfactant and its morphological transformations.</a> J Fluor Chem. 145 (0), 141-147 (2013).</li>
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</ol>

공개 하는 것이 없다.

스 티 렌 및 PFO의 야누스 방울을 포함 하는 유화 제 수성 SDS 농도의 무게 분 율의 10 wt % 및 스 티 렌과 PFO의 어떤 무게 분수 위에서 얻은 했다. 야누스 방울을 포함 하는 유화 액의 다양 한 무게 분수 30 분 생산 되었습니다가지고 때 모든 구성에 대 한 빈 폴리스 티 렌 입자를 얻을 수 있습니다. 이러한 결과 프로토콜 여기 간단 시연 나타냅니다. 또한, 지름, 구멍 크기, 및 구멍 볼륨 빈 폴리스 티 렌 입자의 중 합 시간 여기 설명 하는 방법 제어할 수 있습니다.
우리는 빈 폴리스 티 렌 입자의 제조에 대 한 다음과 같은 메커니즘을 제안. 80 ° c.에 수성 SDS 솔루션, 스 티 렌, 그리고 PFO의 혼합물을 포함 하는 O/W 유화 액에서 스 티 렌 및 PFO의 구성 된 야누스 기름 방울 형성 PFO 방울 방울 스 티 렌의 표면에 있습니다. KPS, 중 합의 창시자 물 단계에서 수용 성 이기 때문에는 중 합 스틸 렌/수성 SDS 솔루션의 인터페이스에서 진행 됩니다. 삼항 유화에 스 티 렌 PFO, 아니 polymerizable 그룹 데 그 온도에 유화 액에 남아 반면 80 ° C에서 폴리스 티 렌을 생산 했다. 후에 중 합 종료 되었습니다, 폴리스 티 렌 입자에 구멍 PFO의 제거에 의해 형성 되었다. 따라서, 빈 컵 형 폴리스 티 렌 입자 표면에 있는 구멍을 갖는 스 티 렌 및 PFO와 모든 중 합 시간 조작된 atany 무게 분수 했다.
90 분에 대 한 과격 한 중 합에 의해 조작 빈 폴리스 티 렌 입자를 톨루엔의 작은 금액을 추가 빈 폴리스 티 렌 입자에 있는 구멍을 밀봉 하는 것에 대 한 제공 합니다. 이 현상은 혁 외.9에 의해 보고와 일치. Capsulation 메서드를 사용 하면 입자에서는 수성 물자를 포함할 수 있습니다.
이 문서에서는, 우리는 없는 상호 혼합할 수 있는 스 티 렌 및 PFO, 구성 된 야누스 기름 방울을 사용 하 여 투명 고분자 입자의 제조 방법을 증명 하고있다. 비 구형 고분자 입자의 준비 방법에는 이미 다양 한 애플 리 케이 션에 그들의 잠재적인 사용 때문에 연구 되었습니다. 야누스 탄화수소 기름과 탄 화 불 소 기름 기름 방울을 사용 하 여이 전략 단위체 및 약물 전달 시스템의 다양 한 종류에서 비 구형 고분자 입자의 제조에 적용 됩니다.

그것은 잘 알려진 보조 개 입자, 반구, 디스크, 및 ellipsoids, 시드 polymerizations,12, 에 의해 준비 된 구형 고분자 입자 다양 한 산업 신청에서 널리 사용 되었다, 3, microreactor를 사용 하 여 특정 형상4,5, 비 구형 모노 머 방울의 사진 중 합 자기 조직 폴리머6의 재와 구형 고분자의 변형 사용 하 여 기계 외부에 의해 입자 힘7,8. 특히, 마이크로미터 크기 투명 폴리머 입자 용 매9,10 사용 하 여 여러 유화 중 합에 의해 부 어 구형 고분자 입자에서 좋은 용 매 증발에 의해 날조되는 11 , 12.
이 작품에서는, 우리는 탄화수소와 탄 화 불 소 고분자 입자의 제조에 오일의 상호 immiscibility의 사용에 집중 한다. 하이브리드 계면 활성 제 분자에 탄 화 수소 체인, 탄 화 불 소 체인 있다. 이전, 우리는 기존의 계면 활성 제13,,1415에 관찰 하지 하이브리드 계면의 독특한 속성을 보고 있다. 우리 또한 탄화수소 석유, 탄 화 불 소 기름, 그리고 수성 계면 활성 솔루션 상호 혼합할 수 없는16을 사용 하 여 유화를 공부 했다. 그러나, 유화 제17의 매우 몇 가지 연구 있다. 이러한 연구는 탄화수소 오일과 fluorocarbon 수성 계면 활성 솔루션에서 구성 된 기름 작은 물방울의 형태를 설명 했습니다.
여기, 우리가 과격 한 중 합 유화 탄화수소 석유, 탄 화 불 소 기름, 및 수성 나트륨 dodecylsulfate (SDS) 솔루션의 구성에 기름 방울을 사용 하 여 투명 고분자 입자의 제조의 상세한 프로토콜을 보여줍니다. 우리는 비 구형 고분자 입자의 준비에 대 한 기존의 방법에서 다른 새로운 전략 제안. 이 방법은 단순히 짧은 시간에 빈 폴리머 입자를 조작 수 있습니다. 또한, 투명 폴리머 입자를 통해 microcapsules의 준비의 프로토콜 표시 됩니다.

<table><tbody><tr><td>&lt;strong&gt;Material&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Sodium dodecylsulfate, 95.0%</td><td>Wako Pure Chemical Industries, Ltd.</td><td>192-08672</td><td></td></tr><tr><td>Styrene, 99.0%</td><td>Tokyo Chemical Industry Co. Ltd.</td><td>S0095</td><td></td></tr><tr><td>Perfluorooctane, 99%</td><td>Fluorochem Ltd.</td><td>8706</td><td></td></tr><tr><td>Coumarin 102, 97.0%</td><td>Tokyo Chemical Industry Co. Ltd.</td><td>C2267</td><td></td></tr><tr><td>Calcein</td><td>Dojindo Molecular Technologies, Inc.</td><td>C001</td><td></td></tr><tr><td>Potassium peroxodisulfate, 98.0%</td><td>Kanto Chemical Co., Inc.</td><td>32375-30</td><td></td></tr><tr><td>Pyrene, 97.0%</td><td>Wako Pure Chemical Industries, Ltd.</td><td>167-05302</td><td></td></tr><tr><td>Ethanol, 99.5%</td><td>Kanto Chemical Co., Inc.</td><td>14033-00</td><td></td></tr><tr><td>Toluene, 99.5%</td><td>Kanto Chemical Co., Inc.</td><td>40180-00</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Equipment&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Scanning Electron Microscope</td><td>Hitachi High-Technologies Corporation</td><td>S-4800</td><td></td></tr><tr><td>Scanning Transmission Electron Microscope</td><td>Hitachi High-Technologies Corporation</td><td>S-4800</td><td></td></tr><tr><td>Ultrasonic cleaner</td><td>Branson Ultrasonics, Emerson Japan, Ltd.</td><td>Model 3510</td><td></td></tr><tr><td>Centrifuge</td><td>AS ONE Corporation</td><td>CN-1050</td><td></td></tr></tbody></table>

preparation of hollow polystyrene particles and microcapsules by radical polymerization of janus droplets consisting of hydrocarbon and fluorocarbon oils

이 문서에서는, 우리는 증명 하고있다 중공 입자와 탄화수소 기름 (스 티 렌)와 탄 화 불 소 기름 구성 된 기름 방울을 사용 하 여 microcapsules를 생성 하는 방법 (퍼플-n-옥 탄, PFO) 수성 계면 활성 제 (나트륨 dodecylsulfate에 SDS) 솔루션입니다. 탄 화 불 소 오일을 혼합할 수 있는 탄화수소 오일 이므로, 두 개의 오일 구분 됩니다. 유화 액 80 ° c.에 스 티 렌/PFO/수성 SDS 솔루션 혼합물을 교 반에 의해 준비 된다 유화 액의 종류와는 유화 액 작은 물방울의 형태는 가벼운 현미경 및 스캐닝 공초점 형광 현미경으로 관찰 된다. 기름 방울 상호 혼합할 수 없는 스 티 렌 및 PFO의 구성 된 야누스 형 형태학을 수성 SDS 솔루션에서 형성 된다 발견 된다. 폴리스 티 렌 입자는 80 ° c.에 스 티 렌/PFO/수성 SDS 솔루션의 삼항 혼합물의 과격 한 중 합 가공 폴리스 티 렌의 형태학 스캐닝 전자 현미경 및 스캐닝 전송 전자 현미경 관찰에 의해 확인 됩니다. 이러한 관측 표면에 단일 구멍 빈 폴리스 티 렌 입자의 준비를 보여줍니다. 우리의 지식,이 방법은 탄화수소와 탄 화 불 소 오일의 immiscibility를 사용 하 여 새로운 전략 이다. 빈 입자는 또한 microcapsules의 준비에 적용할 수 있습니다.

빛과 공초점 형광 현미경 관찰을 스캔 형태학과 스 티 렌, PFO, 그리고 5mm SDS 수 용액 (그림 1)의 구성 된 유화 액 작은 물방울의 구성을 결정 하기 위해 수행 되었습니다. 스캐닝 전자 현미경 (SEM) 및 스캐닝 전송 전자 현미경 (줄기) 관측 중공 입자와 microcapsules (그림 2)의 형성을 보여주었다.
DIC와 스 티 렌/PFO/SDS 수 용액의 삼항 혼합물의 구성 된 유화 액의 공초점 형광 현미경 이미지 연속 단계 임을 나타냅니다 SDS 수 용액, calcein에 해당 하는 녹색 형광 관찰 되었다 때문에 연속 위상, 따라서, 기름에 물 타입 유화 형성 됩니다 (그림 1a 및 1b). 이러한 이미지는 또한 기름 방울의 스 티 렌 PFO, 혼합할 수 있는 상호 되지 않은 구성 다는 것을 보여준다. DIC와 coumarin 102를 포함 하는 유화 액의 공초점 형광 현미경 이미지 공개 PFO 물방울은 물과 스 티 렌 드롭릿 사이의 인터페이스에 위치. 스 티 렌, PFO, 그리고 5mm 폼 유화 야누스를 포함에 SDS 용액의 삼항 혼합물 오일 방울, 스 티 렌과 PFO의 구성.
STEM 및 SEM 이미지 스 티 렌/PFO/5 m m 수성 SDS 솔루션의 유화 액의 과격 한 중 합에 의해 조작 하는 고분자 입자의 표면에 구멍 빈 비 구형 폴리스 티 렌 입자의 준비를 표시 (그림 2a , 2b). 형성 속도, 직경, 및 빈 폴리스 티 렌 입자의 구멍 크기는 STEM 및 SEM을 통해 200 입자를 관찰 하 여 평가 됩니다. SEM 관찰에서 추정 하는 평균된 직경 1.3 µ m 이다. 크기는 동적 산란 측정에서 예상 하는 폴리머 입자의 직경 일치 합니다. 대형 비율은 약 100% 이다. 평균된 구멍 크기와 빈 입자에서 구멍의 볼륨은 0.8 ± 0.4 µ m 및 0.9 ± 0.4 µ m3, 각각. 따라서, 0.8 µ m 구멍 데 빈 폴리스 티 렌 입자는 유화에 스 티 렌을 포함 하는 야누스 방울의 과격 한 중 합에 의해 조작 했다.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56922/56922fig1.jpg"> 그림 1입니다. 스 티 렌, PFO, 그리고 5mm SDS 수 용액의 구성 된 유화 액 작은 물방울의 형태학. (a) 차동 간섭 콘트라스트 (DIC) 및 포함 하는 수성 형광 calcein 유화 액의 (b) 공초점 형광 현미경 이미지. (c) DIC와 (d) 공초점 형광 현미경의 이미지는 유화 액 기름 녹는 형광 coumarin 102를 포함 하. <a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56922/56922fig1large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭 하십시오.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56922/56922fig2.jpg"> 그림 2입니다. 빈 폴리스 티 렌 입자와 microcapsules 스 티 렌, PFO, 그리고 5mm SDS 수 용액의 구성 된 유화 제를 사용 하 여 날조의 형태학. (a) 빈 폴리스 티 렌 입자의 SEM 및 (b) 줄기 이미지. (c) 전자 현미경 및 (d) 줄기의 이미지는 microcapsules. <a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56922/56922fig2large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭 하십시오.</a>

Watch this Scientific Journal Video about Preparation of Hollow Polystyrene Particles and Microcapsules by Radical Polymerization of Janus Droplets Consisting of Hydrocarbon and Fluorocarbon Oils at J

Preparation of Hollow Polystyrene Particles and Microcapsules by Radical Polymerization of Janus Droplets Consisting of Hydrocarbon and Fluorocarbon Oils

빈 폴리스 티 렌 입자와 탄화수소와 탄 화 불 소 오일을 구성 된 야누스 방울의 과격 한 중 합에 의해 Microcapsules의 준비

투명 폴리머 입자와 과격 한 중 합 스틸 렌, 퍼플-n구성 된 유화 제를 사용 하 여 microcapsules의 제조에 대 한 프로토콜-옥 탄, 그리고 수성 SDS (나트륨 dodecylsulfate) 솔루션 제시.

In this article, we have demonstrated a method for producing hollow particles and microcapsules using oil droplets consisting of hydrocarbon oil (styrene) ...

preparation-hollow-polystyrene-particles-microcapsules-radical

Nanyang Technological University

Research

JoVE Journal

Chemistry

9.9K 조회수.  Tokyo University of Science. 투명 폴리머 입자와 과격 한 중 합 스틸 렌, 퍼플-n구성 된 유화 제를 사용 하 여 microcapsules의 제조에 대 한 프로토콜-옥 탄, 그리고 수성 SDS (나트륨 dodecylsulfate) 솔루션 제시.

비디오: Preparation of Hollow Polystyrene Particles and Microcapsules by Radical Polymerization of Janus Droplets Consisting of Hydrocarbon and Fluorocarbon Oils

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

Using Polystyrene-block-poly(acrylic acid)-coated Metal Nanoparticles as Monomers for Their Homo- and Co-polymerization

We present a template-free method for &#8220;polymerizing&#8221; nanoparticles into long chains without side branches. A variety of nanoparticles are encapsulated in polystyrene-block-poly(acrylic acid) (PSPAA) shells and then used as monomers for their self-assembly. Spherical PSPAA micelles upon acid treatment are known to assemble into cylindrical micelles. Exploiting this tendency, the core-shell nanoparticles are induced to aggregate, coalesce, and then transform into long chains. When more than one type of nanoparticles are used, random and block &#8220;copolymers&#8221; of nanoparticles can be obtained. Detailed procedures are reported for the PSPAA encapsulation of nanoparticles, homo- and co-polymerization of the core-shell nanoparticles, separation and purification of the resulting nanoparticle chains. Transformations of single-line chains into double- and triple-line chains are also presented. The synergy between the polymer shell and the embedded nanoparticles leads to an unusual chain-growth polymerization mode, giving long nanoparticle chains that are distinct from the products of the traditional step-growth aggregation process.

Using Polystyrene-block-polyacrylic acid-coated Metal Nanoparticles as Monomers for Their Homo- and Co-polymerization

We report protocols for &#8220;polymerizing&#8221; various types of polymer-encapsulated metal nanoparticles into long chains of &#8220;homo-&#8220; and &#8220;co-polymers&#8221;.

폴리스타이렌을 사용하여<em&gt; 블록</em&gt; 폴리 (아크릴산) 및 이들의 단독 중합체는 공중합 용 단량체로 피복 된 금속 나노 입자

We present a template-free method for &#8220;polymerizing&#8221; nanoparticles into long chains without side branches. A variety of nanoparticles are ...

High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles

Transdisciplinary approaches involving areas such as material design, nanotechnology, chemistry, and immunology have to be utilized to rationally design efficacious vaccines carriers. Nanoparticle-based platforms can prolong the persistence of vaccine antigens, which could improve vaccine immunogenicity1. Several biodegradable polymers have been studied as vaccine delivery vehicles1; in particular, polyanhydride particles have demonstrated the ability to provide sustained release of stable protein antigens and to activate antigen presenting cells and modulate immune responses2-12.
The molecular design of these vaccine carriers needs to integrate the rational selection of polymer properties as well as the incorporation of appropriate targeting agents. High throughput automated fabrication of targeting ligands and functionalized particles is a powerful tool that will enhance the ability to study a wide range of properties and will lead to the design of reproducible vaccine delivery devices.
The addition of targeting ligands capable of being recognized by specific receptors on immune cells has been shown to modulate and tailor immune responses10,11,13 C-type lectin receptors (CLRs) are pattern recognition receptors (PRRs) that recognize carbohydrates present on the surface of pathogens. The stimulation of immune cells via CLRs allows for enhanced internalization of antigen and subsequent presentation for further T cell activation14,15. Therefore, carbohydrate molecules play an important role in the study of immune responses; however, the use of these biomolecules often suffers from the lack of availability of structurally well-defined and pure carbohydrates. An automation platform based on iterative solution-phase reactions can enable rapid and controlled synthesis of these synthetically challenging molecules using significantly lower building block quantities than traditional solid-phase methods16,17.
Herein we report a protocol for the automated solution-phase synthesis of oligosaccharides such as mannose-based targeting ligands with fluorous solid-phase extraction for intermediate purification. After development of automated methods to make the carbohydrate-based targeting agent, we describe methods for their attachment on the surface of polyanhydride nanoparticles employing an automated robotic set up operated by LabVIEW as previously described10. Surface functionalization with carbohydrates has shown efficacy in targeting CLRs10,11 and increasing the throughput of the fabrication method to unearth the complexities associated with a multi-parametric system will be of great value (Figure 1a).

In this article, a high throughput method is presented for the synthesis of oligosaccharides and their attachment to the surface of polyanhydride nanoparticles for further use in targeting specific receptors on antigen presenting cells.

Polyanhydride의 Nanoparticles의 높은 처리량 탄수화물의 합성과 작용화

Transdisciplinary approaches involving areas such as material design, nanotechnology, chemistry, and immunology have to be utilized to rationally design ...

생체공학

High Throughput Single-cell and Multiple-cell Micro-encapsulation

Microfluidic encapsulation methods have been previously utilized to capture cells in picoliter-scale aqueous, monodisperse drops, providing confinement from a bulk fluid environment with applications in high throughput screening, cytometry, and mass spectrometry. We describe a method to not only encapsulate single cells, but to repeatedly capture a set number of cells (here we demonstrate one- and two-cell encapsulation) to study both isolation and the interactions between cells in groups of controlled sizes. By combining drop generation techniques with cell and particle ordering, we demonstrate controlled encapsulation of cell-sized particles for efficient, continuous encapsulation. Using an aqueous particle suspension and immiscible fluorocarbon oil, we generate aqueous drops in oil with a flow focusing nozzle. The aqueous flow rate is sufficiently high to create ordering of particles which reach the nozzle at integer multiple frequencies of the drop generation frequency, encapsulating a controlled number of cells in each drop. For representative results, 9.9 &mu;m polystyrene particles are used as cell surrogates. This study shows a single-particle encapsulation efficiency Pk=1 of 83.7% and a double-particle encapsulation efficiency Pk=2 of 79.5% as compared to their respective Poisson efficiencies of 39.3% and 33.3%, respectively. The effect of consistent cell and particle concentration is demonstrated to be of major importance for efficient encapsulation, and dripping to jetting transitions are also addressed. 
Introduction 
Continuous media aqueous cell suspensions share a common fluid environment which allows cells to interact in parallel and also homogenizes the effects of specific cells in measurements from the media. High-throughput encapsulation of cells into picoliter-scale drops confines the samples to protect drops from cross-contamination, enable a measure of cellular diversity within samples, prevent dilution of reagents and expressed biomarkers, and amplify signals from bioreactor products. Drops also provide the ability to re-merge drops into larger aqueous samples or with other drops for intercellular signaling studies.1,2 The reduction in dilution implies stronger detection signals for higher accuracy measurements as well as the ability to reduce potentially costly sample and reagent volumes.3 Encapsulation of cells in drops has been utilized to improve detection of protein expression,4 antibodies,5,6 enzymes,7 and metabolic activity8 for high throughput screening, and could be used to improve high throughput cytometry.9 Additional studies present applications in bio-electrospraying of cell containing drops for mass spectrometry10 and targeted surface cell coatings.11 Some applications, however, have been limited by the lack of ability to control the number of cells encapsulated in drops. Here we present a method of ordered encapsulation12 which increases the demonstrated encapsulation efficiencies for one and two cells and may be extrapolated for encapsulation of a larger number of cells.
To achieve monodisperse drop generation, microfluidic "flow focusing" enables the creation of controllable-size drops of one fluid (an aqueous cell mixture) within another (a continuous oil phase) by using a nozzle at which the streams converge.13 For a given nozzle geometry, the drop generation frequency f and drop size can be altered by adjusting oil and aqueous flow rates Qoil and Qaq. As the flow rates increase, the flows may transition from drop generation to unstable jetting of aqueous fluid from the nozzle.14 
When the aqueous solution contains suspended particles, particles become encapsulated and isolated from one another at the nozzle. For drop generation using a randomly distributed aqueous cell suspension, the average fraction of drops Dk containing k cells is dictated by Poisson statistics, where Dk = &lambda;k exp(-&lambda;)/(k!) and &lambda; is the average number of cells per drop. The fraction of cells which end up in the "correctly" encapsulated drops is calculated using Pk = (k x Dk)/&Sigma;(k' x Dk'). The subtle difference between the two metrics is that Dk relates to the utilization of aqueous fluid and the amount of drop sorting that must be completed following encapsulation, and Pk relates to the utilization of the cell sample. As an example, one could use a dilute cell suspension (low &lambda;) to encapsulate drops where most drops containing cells would contain just one cell. While the efficiency metric Pk would be high, the majority of drops would be empty (low Dk), thus requiring a sorting mechanism to remove empty drops, also reducing throughput.15
Combining drop generation with inertial ordering provides the ability to encapsulate drops with more predictable numbers of cells per drop and higher throughputs than random encapsulation. Inertial focusing was first discovered by Segre and Silberberg16 and refers to the tendency of finite-sized particles to migrate to lateral equilibrium positions in channel flow. Inertial ordering refers to the tendency of the particles and cells to passively organize into equally spaced, staggered, constant velocity trains. Both focusing and ordering require sufficiently high flow rates (high Reynolds number) and particle sizes (high Particle Reynolds number).17,18 Here, the Reynolds number Re =uDh/&nu; and particle Reynolds number Rep =Re(a/Dh)2, where u is a characteristic flow velocity, Dh [=2wh/(w+h)] is the hydraulic diameter, &nu; is the kinematic viscosity, a is the particle diameter, w is the channel width, and h is the channel height. Empirically, the length required to achieve fully ordered trains decreases as Re and Rep increase. Note that the high Re and Rep requirements (for this study on the order of 5 and 0.5, respectively) may conflict with the need to keep aqueous flow rates low to avoid jetting at the drop generation nozzle. Additionally, high flow rates lead to higher shear stresses on cells, which are not addressed in this protocol. The previous ordered encapsulation study demonstrated that over 90% of singly encapsulated HL60 cells under similar flow conditions to those in this study maintained cell membrane integrity.12 However, the effect of the magnitude and time scales of shear stresses will need to be carefully considered when extrapolating to different cell types and flow parameters. The overlapping of the cell ordering, drop generation, and cell viability aqueous flow rate constraints provides an ideal operational regime for controlled encapsulation of single and multiple cells.
Because very few studies address inter-particle train spacing,19,20 determining the spacing is most easily done empirically and will depend on channel geometry, flow rate, particle size, and particle concentration. Nonetheless, the equal lateral spacing between trains implies that cells arrive at predictable, consistent time intervals. When drop generation occurs at the same rate at which ordered cells arrive at the nozzle, the cells become encapsulated within the drop in a controlled manner. This technique has been utilized to encapsulate single cells with throughputs on the order of 15 kHz,12 a significant improvement over previous studies reporting encapsulation rates on the order of 60-160 Hz.4,15 In the controlled encapsulation work, over 80% of drops contained one and only one cell, a significant efficiency improvement over Poisson (random) statistics, which predicts less than 40% efficiency on average.12
In previous controlled encapsulation work,12 the average number of particles per drop &lambda; was tuned to provide single-cell encapsulation. We hypothesize that through tuning of flow rates, we can efficiently encapsulate any number of cells per drop when &lambda; is equal or close to the number of desired cells per drop. While single-cell encapsulation is valuable in determining individual cell responses from stimuli, multiple-cell encapsulation provides information relating to the interaction of controlled numbers and types of cells. Here we present a protocol, representative results using polystyrene microspheres, and discussion for controlled encapsulation of multiple cells using a passive inertial ordering channel and drop generation nozzle.

Combining monodisperse drop generation with inertial ordering of cells and particles, we describe a method to encapsulate a desired number of cells or particles in a single drop at kHz rates. We demonstrate efficiencies twice exceeding those of unordered encapsulation for single- and double-particle drops.

높은 처리량 마이크로 캡슐 단일 셀 및 다중 셀

Microfluidic encapsulation methods have been previously utilized to capture cells in picoliter-scale aqueous, monodisperse drops, providing confinement ...

Facile Synthesis of Worm-like Micelles by Visible Light Mediated Dispersion Polymerization Using Photoredox Catalyst

Presented herein is a protocol for the facile synthesis of worm-like micelles by visible light mediated dispersion polymerization. This approach begins with the synthesis of a hydrophilic poly(oligo(ethylene glycol) methyl ether methacrylate) (POEGMA) homopolymer using reversible addition-fragmentation chain-transfer (RAFT) polymerization. Under mild visible light irradiation (&#955; = 460 nm, 0.7 mW/cm2), this macro-chain transfer agent (macro-CTA) in the presence of a ruthenium based photoredox catalyst, Ru(bpy)3Cl2 can be chain extended with a second monomer to form a well-defined block copolymer in a process known as Photoinduced Electron Transfer RAFT (PET-RAFT). When PET-RAFT is used to chain extend POEGMA with benzyl methacrylate (BzMA) in ethanol (EtOH), polymeric nanoparticles with different morphologies are formed in situ according to a polymerization-induced self-assembly (PISA) mechanism. Self-assembly into nanoparticles presenting POEGMA chains at the corona and poly(benzyl methacrylate) (PBzMA) chains in the core occurs in situ due to the growing insolubility of the PBzMA block in ethanol. Interestingly, the formation of highly pure worm-like micelles can be readily monitored by observing the onset of a highly viscous gel in situ due to nanoparticle entanglements occurring during the polymerization. This process thereby allows for a more reproducible synthesis of worm-like micelles simply by monitoring the solution viscosity during the course of the polymerization. In addition, the light stimulus can be intermittently applied in an ON/OFF manner demonstrating temporal control over the nanoparticle morphology.

This article describes a process for producing polymeric self-assembled nanoparticles using visible light mediated dispersion polymerization. Using low energy visible light to control the polymerization allows for the reproducible formation of self-assembled worm-like micelles at high solids content.

Photoredox 촉매를 사용하여 가시 광선 중재 분산 중합에 의해 웜 형상 미셀 손쉬운 합성

Presented herein is a protocol for the facile synthesis of worm-like micelles by visible light mediated dispersion polymerization. This approach begins ...

Anionic Polymerization of an Amphiphilic Copolymer for Preparation of Block Copolymer Micelles Stabilized by &#960;-&#960; Stacking Interactions

In this study, an amphiphilic copolymer that includes a core-forming block with phenyl groups was synthesized by living anionic polymerization of phenyl glycidyl ether (PheGE) on methoxy-polyethylene glycol (mPEG-b-PPheGE). Characterization of the copolymer revealed a narrow molecular distribution (PDI &#60; 1.03) and confirmed the degree of polymerization of mPEG122-b-(PheGE)15. The critical micelle concentration of the copolymer was evaluated using an established fluorescence method with the aggregation behavior evaluated by dynamic light scattering and transmission electronic microscopy. The potential of the copolymer for use in drug delivery applications was evaluated in a preliminary manner including in vitro biocompatibility, loading and release of the hydrophobic anti-cancer drug doxorubicin (DOX). A stable micelle formulation of DOX was prepared with drug loading levels up to 14% (wt%), drug loading efficiencies &#62; 60% (w/w) and sustained release of drug over 4 days under physiologically relevant conditions (acidic and neutral pH, presence of albumin). The high drug loading level and sustained release is attributed to stabilizing &#960;-&#960; interactions between DOX and the core-forming block of the micelles.

The key steps of living anionic polymerization of phenyl glycidyl ether (PheGE) on methoxy-polyethylene glycol (mPEG-b-PPheGE) are described. The resulting block copolymer micelles (BCMs) were loaded with doxorubicin 14% (wt%) and sustained release of drug over 4 days under physiologically relevant conditions was obtained.

π-π 스태킹 상호 작용에 의해 안정화 된 블록 공중 합체 미셀 준비하는 양친 매성 공중 합체의 음이온 중합

In this study, an amphiphilic copolymer that includes a core-forming block with phenyl groups was synthesized by living anionic polymerization of phenyl ...

Fabrication of Polymer Microspheres for Optical Resonator and Laser Applications

This paper describes three methods of preparing fluorescent microspheres comprising &#960;-conjugated or non-conjugated polymers: vapor diffusion, interface precipitation, and mini-emulsion. In all methods, well-defined, micrometer-sized spheres are obtained from a self-assembling process in solution. The vapor diffusion method can result in spheres with the highest sphericity and surface smoothness, yet the types of the polymers able to form these spheres are limited. On the other hand, in the mini-emulsion method, microspheres can be made from various types of polymers, even from highly crystalline polymers with coplanar, &#960;-conjugated backbones. The photoluminescent (PL) properties from single isolated microspheres are unusual: the PL is confined inside the spheres, propagates at the circumference of the spheres via the total internal reflection at the polymer/air interface, and self-interferes to show sharp and periodic resonant PL lines. These resonating modes are so-called "whispering gallery modes" (WGMs). This work demonstrates how to measure WGM PL from single isolated spheres using the micro-photoluminescence (&#181;-PL) technique. In this technique, a focused laser beam irradiates a single microsphere, and the luminescence is detected by a spectrometer. A micromanipulation technique is then used to connect the microspheres one by one and to demonstrate the intersphere PL propagation and color conversion from coupled microspheres upon excitation at the perimeter of one sphere and detection of PL from the other microsphere. These techniques, &#181;-PL and micromanipulation, are useful for experiments on micro-optic application using polymer materials.

Protocols for the synthesis of microspheres from polymers, the manipulation of microspheres, and micro-photoluminescence measurements are presented.

광 공진기 및 레이저 응용을위한 고분자 미소 구의 제조

This paper describes three methods of preparing fluorescent microspheres comprising &#960;-conjugated or non-conjugated polymers: vapor diffusion, ...

공학

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 ...

Synthesis of Monodisperse Cylindrical Nanoparticles via Crystallization-driven Self-assembly of Biodegradable Block Copolymers

The production of monodisperse cylindrical micelles is a significant challenge in polymer chemistry. Most cylindrical constructs formed from diblock copolymers are produced by one of three techniques: thin film rehydration, solvent switching or polymerization-induced self-assembly, and produce only flexible, polydisperse cylinders. Crystallization-driven self-assembly (CDSA) is a method which can produce cylinders with these properties, by stabilizing structures of a lower curvature due to the formation of a crystalline core. However, the living polymerization techniques by which most core-forming blocks are formed are not trivial processes and the CDSA process may yield unsatisfactory results if carried out incorrectly. Here, the synthesis of cylindrical nanoparticles from simple reagents is shown. The drying and purification of reagents prior to a ring-opening polymerization of &#949;-caprolactone catalyzed by diphenyl phosphate is described. This polymer is then chain extended by methyl methacrylate (MMA) followed by N,N-dimethyl acrylamide (DMA) using reversible addition&#8722;fragmentation chain-transfer (RAFT) polymerization, affording a triblock copolymer that can undergo CDSA in ethanol. The living CDSA process is outlined, the results of which yield cylindrical nanoparticles up to 500 nm in length and a length dispersity as low as 1.05. It is anticipated that these protocols will allow others to produce cylindrical nanostructures and elevate the field of CDSA in the future.

Crystallization-driven self-assembly (CDSA) displays the unique ability to fabricate cylindrical nanostructures of narrow length distributions. The organocatalyzed ring-opening polymerization of &#949;-caprolactone and subsequent chain extensions of methyl methacrylate and N,N-dimethyl acrylamide are demonstrated. A living CDSA protocol that produces monodisperse cylinders up to 500 nm in length is outlined.

생분해성 블록 공합체의 결정화 구동 자가 조립을 통해 단분산 원통형 나노 입자의 합성

The production of monodisperse cylindrical micelles is a significant challenge in polymer chemistry. Most cylindrical constructs formed from diblock ...

A Droplet-Based Microfluidic Approach and Microsphere-PCR Amplification for Single-Stranded DNA Amplicons

Droplet-based microfluidics enable the reliable production of homogeneous microspheres in the microfluidic channel, providing controlled size and morphology of the obtained microsphere. A microsphere copolymerized with an acrydite-DNA probe was successfully fabricated. Different methods such as asymmetric PCR, exonuclease digestion, and isolation on streptavidin-coated magnetic beads can be used to synthesize single-stranded DNA (ssDNA). However, these methods cannot efficiently use large amounts of highly purified ssDNA. Here, we describe a microsphere-PCR protocol detailing how ssDNA can be efficiently amplified and separated from dsDNA simply by pipetting from a PCR reaction tube. The amplification of ssDNA can be applied as potential reagents for the DNA microarray and DNA-SELEX (Systematic evolution of ligands by exponential enrichment) processes.

This work provides a method for the fabrication of droplet-based microfluidic platforms and the application of polyacrylamide microspheres for microsphere-PCR amplification. The microsphere-PCR method makes it possible to obtain single-stranded DNA amplicons without separating double-stranded DNA.

빈 폴리스 티 렌 입자와 탄화수소와 탄 화 불 소 오일을 구성 된 야누스 방울의 과격 한 중 합에 의해 Microcapsules의 준비

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