Bu eser (Grant No finansman Shenzhen temel araştırma tarafından desteklenmiştir JCYJ 20150630170146829, JCYJ20160531195439665 ve JCYJ20160317152359560). Yazarlar, Shenzhen ileri teknoloji enstitüleri, Çin için çekmek Bilimler Akademisi Prof. Y. Chen teşekkür etmek istiyorum.

1. kalıp imalat
<ol><li>İki photomasks bir çizim yazılımı kullanarak tasarlayın. Microchannel yapısı çerçevesinde tarif ve iki ayrı katman maskesi 1 ve 2 aynı çizim dosyası kullanmak, yani farklı kanalları arasındaki tüm bağlantıları sağlamak. Bağımsız bir satıcı 1 µm çözünürlükle tarafından cam tabakta krom için farklı katmanları yazdır. Photomasks negatif polarite ile şeffaf tasarlanmış yapılar, karanlık olduğundan emin olun. Not: Maske 1 dağınık faz giriş kanalı ve bir delik içerir. Maske 2 sürekli faz giriş kanalı, filtre ve çıkış içerir.</li>
	<li>Belirlenen fotolitografi laboratuvarda, 3-inç çapında silikon gofret temiz. Gofret bir spin coater yerleştirin, gofret spin Chuck eklemesi vakum açın. Spin-kat 2-3 mL SU-8 2025 negatif fotorezist 10 için gofret üzerine 1000 rpm, o zaman 30 s s 3000 devirde 20 µm ilk katman kalınlığı sağlamak.</li>
	<li>Yumuşak fırında 6 min için 95 ° C ocağın üzerinde. Oda sıcaklığında (RT) kaplı gofret soğur sonra altında bir collimated 15 mW/cm2, 365 1 maske üzerinden maruz kalmaktadır UV 18 s. sonrası maruz kalma, nm fırında 6 min için 95 ° C ocağın üzerinde sonra gofret için RT. soğutmak izin</li>
	<li>Spin-kaplama işlemi yineleyin. 2-3 mL SU-8 2100 negatif fotorezist 10 için gofret üzerine uygulamak 1000 rpm, o zaman 30 s s 2000 devirde 130 µm ikinci katman kalınlığı sağlamak. 35 dk için 95 ° C ocağın üzerinde yumuşak fırında sonra maske 2 30 için ortaya çıkarmak için ikinci katman fotorezist yer dağınık faz kanal katmanla UV aligner tarafından uyumlu s. Sonrası pozlama, fırında 7 min için 95 ° C ocağın üzerinde.</li>
	<li>Gofret özellikleri üzerinde gofret ortaya çıkıyor, sonra etil alkol ile yıkayın kadar Propilen Glikol Metil Eter asetat 50 mL karıştırılmış bir banyoda çeker tarafından geliştirmek. Son olarak, gofret zor-2s için pişirme için termostatik platform üzerine yerleştirin.</li>
</ol>2. yarı-3D akışı odaklanarak mikrosıvısal çip imalat
<ol><li>Mix PDMS monomer ve üst ve alt katmanları, genellikle 10:1 üst tabaka için ve 8:1 alt tabaka için biraz farklı bir manevra kabiliyetine içinde kür Aracısı otomatik merhem karıştırıcı için 4 dk kullanarak. Not: Üst ve alt PDMS döşeme biraz farklı bir oran içinde hazırlanır (10:1 ve 8:1 sırasıyla) PDMS tabanına kür Ajan, bağ gücü arttırmak. 5:1 oranı için alt katmanı seçildiğinde, daha sert alt PDMS levha hizalamayı zorlaştırır ve esneklik yetersizliği nedeniyle bağ gücü azaltır. Buna ek olarak, alt PDMS levha kalınlığı mikroskop çalışma mesafesi uyum sağlamak için yaklaşık 1 mm olduğunu. 15:1 oranı seçildiğinde çip kolayca yüksek basınç altında deforms.</li>
	<li>90 mm petri kabına tamamlanan silikon kalıp içine karışımı dökün ve 2 kalınlığı sağlamak ~ 3 mm. bir vakum odasında yerleştirin ve kadar tüm hava kabarcıkları ortadan degas. 80 ° c fırında 1 saat için tedavi.</li>
	<li>PDMS RT. kullanmak için en az 3 mm Özellikleri uzak cihazını ve yavaş yavaş silikon gofret PDMS katmandan kapalı soyma bir neşter soğutmak izin. Dağınık faz giriş, sürekli faz giriş ve çıkış bir 0,75 mm çap yumruk kullanarak üst PDMS katmanındaki bir yumruk.</li>
	<li>Toz parçacıkları kaldırmak için PDMS yapışkan bant ile temizleyin. Plazma tedavi alt ve üst PDMS katmanları aynı anda 2 min için bir 300 W plazma temizleyici. Üst katmanı veya alt katmanı çekilebilecek ve özellikleri ile stereo mikroskop ile ilgilenen hizalanır kadar yüzeyleri nispeten kaydırın.</li>
	<li>Aygıtın gücünü artırmak ve bağ tamamlamak bir günlük 120 ° c fırında tedavi.</li>
</ol>3. reaktifler hazırlık
<ol><li>Sürekli aşama çözüm hazırlamak: 18 vol % silikon tabanlı Nonyonik yüzey aktif eriterek tarafından Hexadecane.</li>
	<li>Dağınık faz çözüm hazırlamak: hidrofilik poli (etilen glikol) diacrylate (PEGDA, MW 255) içeren 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone (%98, MW 224) 5 mg/mL konsantrasyonu fotoğraf-Başlatıcı ve rodamine B (olarak, % 95 MW 479.01) adlı bir konsantrasyon 1 mmol/L floresan boyalar olarak.</li>
	<li>1 mL rezervuar pnömatik basınç denetleyicisinin sürekli aşama ile doldurun. 200 µL jel yükleme ipucu dağınık faz ile doldurun.</li>
</ol>4. Sistem Hazırlama
<ol><li>Yarı-3D mikrosıvısal aygıt ile yüksek hızlı bir fotoğraf makinesi ile ilgilenen bir ters optik mikroskobu sahne yerleştirin.</li>
	<li>Fluorlu etilen propilen (FEP) tüp sürekli aşama delikli deliğe kısa bir paslanmaz çelik tüp takarak bağlayın ve dağınık faz delikli deliğe jel yükleme bahşiş ucunu bağlayın. FEP tüp 20 cm uzunluğunda cihazı prize takın ve sonunda bir 15 mL santrifüj tüpü yerleştirin. Not: Sistem yapılandırmasını Şekil 1' de gösterilmiştir.</li>
</ol>5. damlacıkları oluşumu
<ol><li>Cihazın ters bir mikroskop tezgah üzerinde yerleştirin ve farklı kanalları kavşak kabaca mikroskop ışık kaynağı bulunduğu yer alır emin olun. İki aşama kavşak içeren bir bölge, ağız bölgesi ve aşağı akım kanal ters mikroskobunun odaklan.</li>
	<li>Basınç iki aşamadan sıvı yavaş yavaş dağınık aşaması için 15 mbar ve sürekli aşama için 30 mbar ile kesişen bölge teslim etmek için pnömatik basınç denetleyicisi kullanarak ayarlayın. İstikrar ve denge kadar hiçbir kabarcıklar ve PDMS kalıntı taşıyan istikrarlı sıvı akış için 3 dakika bekleyin.</li>
	<li>Giriş parametreleri kontrol yazılımı Kullanıcı arabiriminde. Örneğin dağınık faz baskısı bir sabit sisteme taban düzeyi baskısı olarak ayarlayın, 45 mbar koru. Emülsiyon ayrılık modu için ipucu akış modu jeti üzerinden değiştirilinceye kadar sürekli aşama basınç artışı, sonra istikrar için 5 dakika bekleyin.</li>
	<li>Çıkış deliği damlacıkları toplamak için santrifüj tüpü bağlanma FEP tüp sonuna yerleştirin.</li>
</ol>6. PEGDA parçacıklar toplama ve karakterizasyonu
<ol><li>Santrifüj tüpü aslant fikstür içinde düzeltmek. Sıvı faz akış tüp hızlı katılaşma aracılığıyla PEGDA parçacıkların UV Işınlarına maruz kalma tarafından elde.</li>
	<li>Toplama işlemi tamamlanırken, örnek ve sırasıyla PEGDA parçacıkları ile 20 X ve 60 X nesneleri ile floresan mikroskop gözlemlemek. Not. Floresan fotoğraflar kamera tarafından çekilen bir ısmarlama yazılım yordamı tarafından incelenir. İlk deconvolution R-L algoritması17 out-of-odaklanan ışık ve görüntüdeki nesnelerin açıkgöz kenar algılama yöntemiyle alan ayıklanır küre yitirmesi etkisini ortadan kaldırmak için temel görüntüsüdür; son olarak, her küre nesne çapını Hough dönüşüm18kullanılarak hesaplanabilir. Sonuç olarak, ortalama ve standart sapma küre nesneleri her görüntüde çapları, tahmin edilebilir.</li>
</ol>

<ol>
	<li>Teh, S. Y., Lin, R., Hung, L. H., Lee, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Droplet+microfluidics.">Droplet microfluidics.</a> Lab on a chip. 8, 198-220 (2008).</li><li>Xiao, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+multifunctional+NaYF4:Er3+,Yb3+/PEGDA+hybrid+microspheres:+NIR-light-activated+photopolymerization+and+drug+delivery.">Novel multifunctional NaYF4:Er3+,Yb3+/PEGDA hybrid microspheres: NIR-light-activated photopolymerization and drug delivery.</a> Chemical communications. 49, 1527-1529 (2013).</li><li>Pan, M., Kim, M., Blauch, L., Tang, S. K. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface-functionalizable+amphiphilic+nanoparticles+for+pickering+emulsions+with+designer+fluid-fluid+interfaces.">Surface-functionalizable amphiphilic nanoparticles for pickering emulsions with designer fluid-fluid interfaces.</a> RSC Adv. 6, 39926-39932 (2016).</li><li>Zhang, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+synthesis+of+rigid+nanovesicles+for+hydrophilic+reagents+delivery.">Microfluidic synthesis of rigid nanovesicles for hydrophilic reagents delivery.</a> Angewandte Chemie. 54, 3952-3956 (2015).</li><li>Shembekar, N., Chaipan, C., Utharala, R., Merten, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Droplet-based+microfluidics+in+drug+discovery,+transcriptomics+and+high-throughput+molecular+genetics.">Droplet-based microfluidics in drug discovery, transcriptomics and high-throughput molecular genetics.</a> Lab on a chip. 16, 1314-1331 (2016).</li><li>Soh, G. Y., Yeoh, G. H., Timchenko, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Numerical+investigation+on+the+velocity+fields+during+droplet+formation+in+a+microfluidic+T-junction.">Numerical investigation on the velocity fields during droplet formation in a microfluidic T-junction.</a> Chemical Engineering Science. 139, 99-108 (2016).</li><li>Chiarello, E., Derzsi, L., Pierno, M., Mistura, G., Piccin, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+Oil+Droplets+in+a+Non-Newtonian+Liquid+Using+a+Microfluidic+T-Junction.">Generation of Oil Droplets in a Non-Newtonian Liquid Using a Microfluidic T-Junction.</a> Micromachines. 6, 1825-1835 (2015).</li><li>Moyle, T. M., Walker, L. M., Anna, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlling+thread+formation+during+tipstreaming+through+an+active+feedback+control+loop.">Controlling thread formation during tipstreaming through an active feedback control loop.</a> Lab on a chip. 13, 4534-4541 (2013).</li><li>Moon, B. U., Abbasi, N., Jones, S. G., Hwang, D. K., Tsai, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Water-in-Water+Droplets+by+Passive+Microfluidic+Flow+Focusing.">Water-in-Water Droplets by Passive Microfluidic Flow Focusing.</a> Analytical chemistry. 88, 3982-3989 (2016).</li><li>Zhu, P., Tang, X., Wang, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Droplet+generation+in+co-flow+microfluidic+channels+with+vibration.">Droplet generation in co-flow microfluidic channels with vibration.</a> Microfluidics and Nanofluidics. , 20 (2016).</li><li>Kim, S. H., Kim, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+formation+of+double-emulsion+drops+in+sudden+expansion+channels.">Controlled formation of double-emulsion drops in sudden expansion channels.</a> Journal of colloid and interface science. 415, 26-31 (2014).</li><li>Jeong, W. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+generation+of+submicron+emulsion+droplets+via+highly+stable+tip-streaming+mode+in+microfluidic+devices.">Controlled generation of submicron emulsion droplets via highly stable tip-streaming mode in microfluidic devices.</a> Lab on a chip. 12, 1446-1453 (2012).</li><li>Peng, L., Yang, M., Guo, S. S., Liu, W., Zhao, X. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+interfacial+tension+on+droplet+formation+in+flow-focusing+microfluidic+device.">The effect of interfacial tension on droplet formation in flow-focusing microfluidic device.</a> Biomedical microdevices. 13, 559-564 (2011).</li><li>Nunes, J. K., Tsai, S. S., Wan, J., Stone, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dripping+and+jetting+in+microfluidic+multiphase+flows+applied+to+particle+and+fiber+synthesis.">Dripping and jetting in microfluidic multiphase flows applied to particle and fiber synthesis.</a> J Phys D Appl Phys. , 46 (2013).</li><li>Anna, S. L., Mayer, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microscale+tipstreaming+in+a+microfluidic+flow+focusing+device.">Microscale tipstreaming in a microfluidic flow focusing device.</a> Physics of Fluids. 18, 121512 (2006).</li><li>Liu, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+synthesis+of+QD-encoded+PEGDA+microspheres+for+suspension+assay.">Microfluidic synthesis of QD-encoded PEGDA microspheres for suspension assay.</a> J. Mater. Chem. B. 4, 482-488 (2016).</li><li>Richardson, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bayesian-based+iterative+method+of+image+restoration.">Bayesian-based iterative method of image restoration.</a> Journal of the Optical Society of America. 62, 55-59 (1972).</li><li>Mukhopadhyay, P., Chaudhuri, B. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+survey+of+Hough+Transform.">A survey of Hough Transform.</a> Pattern Recognition. 48, 993-1010 (2015).</li><li>Ward, T., Faivre, M., Stone, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drop+production+and+tip-streaming+phenomenon+in+a+microfluidic+flow-focusing+device+via+an+interfacial+chemical+reaction.">Drop production and tip-streaming phenomenon in a microfluidic flow-focusing device via an interfacial chemical reaction.</a> Langmuir: the ACS journal of surfaces and colloids. 26, 9233-9239 (2010).</li><li>Anna, S. L., Bontoux, N., Stone, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formation+of+dispersions+using+"flow+focusing"+in+microchannels.">Formation of dispersions using "flow focusing" in microchannels.</a> Applied Physics Letters. 82, 364 (2003).</li><li>Kim, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+production+of+emulsion+drops+using+an+electric+field+in+a+flow-focusing+microfluidic+device.">Controlled production of emulsion drops using an electric field in a flow-focusing microfluidic device.</a> Applied Physics Letters. 91, 133106 (2007).</li><li>Dangla, R., Gallaire, F., Baroud, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microchannel+deformations+due+to+solvent-induced+PDMS+swelling.">Microchannel deformations due to solvent-induced PDMS swelling.</a> Lab on a chip. 10, 2972-2978 (2010).</li><li>Chiu, Y. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Universally+applicable+three-dimensional+hydrodynamic+microfluidic+flow+focusing.">Universally applicable three-dimensional hydrodynamic microfluidic flow focusing.</a> Lab on a chip. 13, 1803-1809 (2013).</li><li>Oh, K. W., Lee, K., Ahn, B., Furlani, E. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+of+pressure-driven+microfluidic+networks+using+electric+circuit+analogy.">Design of pressure-driven microfluidic networks using electric circuit analogy.</a> Lab on a chip. 12, 515-545 (2012).</li></ol>

Yazarlar ifşa gerek yok.

Damlacıkları 2D ve yarı-3D mikrosıvısal aygıt kullanarak akış odaklama modunda nesil daha önce çeşitli raporlar8,9,15,19,20geliştirdi, 21. Bu sistemler değil katılaşmış sulu sıvı sodyum sulu bir çözüm deiyonize su8,15,<sup class="xref...

Mikrosıvısal damlacık tabanlı sistemleri son derece mikrometre çapında aralığı1 nanometre üzerinden monodisperse damlacıkları üretmek ve yüksek üretilen iş ilaç bulma2' de, biomolecules3 sentezi büyük potansiyele sahip yeteneği var ,4ve5test tanı. Büyük yüzey alanı hacim oranı ve örneği, bir kaç microliters tüketen ile büyük ölçekli uygulamalar gibi daha küçük damlacıklar benzersiz avantajları nedeniyle teknoloji alanları geniş bir yelpazesi geniş ilgi çekti. İki immiscible sıvı Emülsifikasyon damlacık oluşturmak için en çok kullanılan yöntemlerinden biridir. Alanın önceki raporlarda araştırmacılar farklı damlacık oluşumu geometrileri, akışı odaklanarak ve geometriler Co akan paraleldir, dahil olmak üzere çeşitli geliştirdik. Paraleldir geometride dağınık faz sürekli aşama6,7akar ana kanal dikey bir kanal üzerinden teslim edilir. Tipik iki boyutlu (2D) akışı odaklanarak8,9 geometri, dağınık faz akışı lateral güdülmesini; ve böylece dağınık faz akışı üzerinden nın yaşadığı ortak akan geometri10,11için öte yandan, dağınık Faz akımı tanıtan bir kılcal co-eksenel Co akan geometri için daha büyük bir kılcal damar içine yerleştirilir her yöne.
Damlacık boyutu kanal boyutu ve akış hızı oranı ayarlayarak denetlenir ve ortak akan veya paraleldir tarafından üretilen en küçük boyut mikrometre düzinelerce için sınırlıdır. Akışı odaklanarak damlacık oluşumu sistem için damlacık ayrılık üç modları formu iki aşamalı basınç oranını ayarlayarak ve yüzey aktif konsantrasyon damlama dahil olmak üzere, rejim, jeti rejimi ve15ipucu akarsu. İpucu akış ayrıca iş parçacığı oluşumu ve ince dağınık Faz akımı koni ucundan çizim iplik gözlenen görünümünü adı verilir. Az önceki çalışmalarda damlacıkları göstermiştir birkaç mikrometre ipucu akış rağmen oluşturulabilir 2D veya yarı-3D akışı odaklanarak aygıt8,12işlem. PEGDA çok düşük bir konsantrasyon içeren sulu bir çözüm dağınık faz kullanılan ancak, büzülme oranı PEGDA parçacıkların özgün damlacıkların çapı yaklaşık % 60 fotoğraf-polimerizasyon, seyreltme olmadan PEGDA süre sonra olduğu dağınık faz kararsız ipucu akış modu12' ye götürdü. İnterfacial gerilim olduğunu emülsiyon sürecinin önemli bir parametre ve yüzey aktif ek damlacık boyutu, daha yüksek üretimi frekans13, son derece eğimli uç, azaltmak için önde gelen sürekli aşama sıvı içine nedeniyle azalır ve istikrarsızlık14engelliyor. Ayrıca, toplu yüzey aktif konsantrasyonu kritik micelle konsantrasyonu çok yüksek olduğunda, interfacial gerilim doymuş devlet13 ' te yaklaşık değişmez ve ipucu akış modu15oluşabilir.
Bu kağıt, yukarıdaki gözlemleri dayalı çok katmanlı yumuşak taş baskı yöntemi ile fabrikasyon bir yarı-3D akışı odaklanarak mikrosıvısal aygıtı kullanarak PEGDA damlacıkları üretimi için facile bir yaklaşım geliştirdik. Böylece dağınık faz üzerinden yukarı ve aşağı lateral nın yaşadığı farklı tipik 2D aygıt akışı odaklanarak, yarı-3D akışı odaklanarak cihaz sığ dağınık faz kanal ve derin sürekli aşama kanal vardır. Bu akış odaklama modu için enerji ve damlacık ayrılık için gerekli basıncı azaltarak daha geniş ayar aralığı sağlar. Önceki rapor12, dağınık faz saf PEGDAcontaining fotoğraf-Başlatıcı, PEGDA parçacıklar büzülme oranı % 1016' dan daha düşük olduğundan emin farklıdır; ve sürekli aşama silikon tabanlı Nonyonik yüzey aktif yüksek toplu konsantrasyon ile eriterek hexadecane karışımıdır. Boyutu kontrol edilebilir ve düzgün damlacıkları iki aşamalı basınç oranını ayarlayarak üretildi. Damlacık ayrılık ipucu akış modu için jeti mod değişiklikleri işlerken damlacıkların çapı 80 µm için 1 µm değiştirir. Buna ek olarak, PEGDA parçacık UV Işınlarına maruz kalma altında fotoğraf-polimerizasyon sürecinde sentez. Damlacık üretimi mikrosıvısal sistem imalat kolaylığı ile biyolojik uygulamalar için daha fazla olanak sağlar.

<table><tbody><tr><td>Silicon wafer</td><td>Huashi Co., Ltd</td><td></td><td></td></tr><tr><td>SU-8 2025, 2100</td><td>Microchem Co.</td><td>Y111069</td><td></td></tr><tr><td>SU-8 developer</td><td>Microchem Co.</td><td>Y020100</td><td></td></tr><tr><td>Chromium mask</td><td>Qingyi Precision Mask Making Co., Ltd</td><td></td><td></td></tr><tr><td>polydimethylsiloxane(PDMS)</td><td>Dow Corning</td><td>Sylgard 184</td><td></td></tr><tr><td>poly(ethylene glycol) diacrylate (PEGDA)</td><td>Sigma</td><td>26570-48-9</td><td></td></tr><tr><td>2-hydroxy-40-(2-hydroxyethoxy)-2-methylpropiophenone</td><td>TCI</td><td>H1361-5G</td><td>photoinitiator</td></tr><tr><td>Hexadecane</td><td>Sigma</td><td>544-76- 3</td><td></td></tr><tr><td>ABIL EM 90</td><td>CHT</td><td>144243-53-8</td><td>surfactants</td></tr><tr><td>Rhodamine B</td><td>Aladdin</td><td>81-88-9</td><td>fluorescent dye</td></tr><tr><td>Spin Coater</td><td></td><td></td><td></td></tr><tr><td>Lithography machine</td><td></td><td></td><td></td></tr><tr><td>Automatic ointment agitator</td><td>Thinky</td><td>ARV-310</td><td></td></tr><tr><td>Oven</td><td>BluePard</td><td></td><td></td></tr><tr><td>Optical microscope</td><td>OLYMPUS</td><td>IX71</td><td></td></tr><tr><td>High-speed camera</td><td>Hamamatsu, Japan</td><td>ORCA-flash</td><td></td></tr><tr><td>MAESFLO Microfluidic Fluid Control System</td><td>FLUIGENT</td><td>MFCS-EZ</td><td></td></tr><tr><td>UV lamp</td><td>FUTANSI</td><td></td><td>365 nm UV light, 8000 MW/CM&lt;sup&gt;2&lt;/sup&gt;</td></tr></tbody></table>

generation of size-controlled poly (ethylene glycol) diacrylate droplets via semi-3-dimensional flow focusing microfluidic devices

Üniforma ve boyut kontrol edilebilir poli (etilen glikol) diacrylate (PEGDA) damlacıkları işlemi bir mikrosıvısal cihazın odaklama akış yoluyla yapılabilir. Bu kağıt bir semi-three-dimensional (yarı-3D) akışı odaklanarak mikrosıvısal çip damlacık oluşumu için öneriyor. Polydimethylsiloxane (PDMS) çip çok katmanlı yumuşak taş baskı yöntemiyle fabrikasyon. Yüzey aktif içeren Hexadecane sürekli aşama kullanıldı ve PEGDA ultraviyole (UV) fotoğraf-başlatıcı ile dağınık dönemdi. Yüzey bırakmak yerel yüzey gerilimi izin ve küçük mikro damlacıkları kırarak terfi daha cusped bahşiş kurdu. Dağınık faz baskısı sürekli damlacıkları boyutu ile artan sürekli aşama basınç dağınık Faz akımı kırılmış önce daha küçük hale geldi. Sonuç olarak, damlacıkları boyutu çeşitlemesiyle 1 µm den çapı 80 µm seçici basınç oranı iki giriş kanalları değiştirerek elde edilebilir ve varyasyon ortalama katsayısı % 7'in altına olarak tahmin edildi. Ayrıca, damlacıkları fotoğraf-polimerizasyon için UV Işınlarına maruz kalma tarafından mikro-boncuk çevirebilir. Böyle mikro-boncuk yüzeyi conjugating biomolecules biyoloji ve Kimya alanlarında birçok potansiyel uygulamalar var.

Yarı-3D akışı odaklanarak mikrosıvısal çip yukarıda açıklandığı gibi çok katmanlı yumuşak litografi teknikleri kullanarak fabrikasyon. İmalat süreci ve sonuçları elde etmek için Ana kalıp Şekil 2' de gösterilen protocolare. 65 µm geniş kanal dağınık faz ve bir 50 µm geniş orifis (Şekil 2bir) tanıtmak için sağlar, ilk katman 20 µm kalınlığında olduğunu. Bir ek 130 µm kalı...

Watch this Scientific Journal Video about Generation of Size-controlled Poly ethylene Glycol Diacrylate Droplets via Semi-3-Dimensional Flow Focusing Microfluidic Devices at JoVE.com

Generation of Size-controlled Poly ethylene Glycol Diacrylate Droplets via Semi-3-Dimensional Flow Focusing Microfluidic Devices

Burada, imalat süreçleri ve bir semi-three-dimensional (yarı-3D) akışı odaklanarak mikrosıvısal çip damlacık oluşumu için doğrulama deneyler göstermek için bir iletişim kuralı mevcut.

Poli (etilen glikol) Diacrylate damlacıklar yoluyla yarı-3 boyutlu akış odaklanarak mikrosıvısal cihazlar boyutu kontrol nesil

Uniform and size-controllable poly (ethylene glycol) diacrylate (PEGDA) droplets could be produced via the flow focusing process in a microfluidic device. ...

generation-size-controlled-poly-ethylene-glycol-diacrylate-droplets

Universidad San Sebastian

Research

JoVE Journal

Engineering

Generation of Size-controlled Poly (ethylene Glycol) Diacrylate Droplets via Semi-3-Dimensional Flow Focusing Microfluidic Devices

7.7K Görüntüleme.  Tsinghua University. Burada, imalat süreçleri ve bir semi-three-dimensional (yarı-3D) akışı odaklanarak mikrosıvısal çip damlacık oluşumu için doğrulama deneyler göstermek için bir iletişim kuralı mevcut.

Video: Generation of Size-controlled Poly ethylene Glycol Diacrylate Droplets via Semi-3-Dimensional Flow Focusing Microfluidic Devices

Fluorescence detection methods for microfluidic droplet platforms

The development of microfluidic platforms for performing chemistry and biology has in large part been driven by a range of potential benefits that accompany system miniaturisation. Advantages include the ability to efficiently process nano- to femoto- liter volumes of sample, facile integration of functional components, an intrinsic predisposition towards large-scale multiplexing, enhanced analytical throughput, improved control and reduced instrumental footprints.1
In recent years much interest has focussed on the development of droplet-based (or segmented flow) microfluidic systems and their potential as platforms in high-throughput experimentation.2-4 Here water-in-oil emulsions are made to spontaneously form in microfluidic channels as a result of capillary instabilities between the two immiscible phases. Importantly, microdroplets of precisely defined volumes and compositions can be generated at frequencies of several kHz. Furthermore, by encapsulating reagents of interest within isolated compartments separated by a continuous immiscible phase, both sample cross-talk and dispersion (diffusion- and Taylor-based) can be eliminated, which leads to minimal cross-contamination and the ability to time analytical processes with great accuracy. Additionally, since there is no contact between the contents of the droplets and the channel walls (which are wetted by the continuous phase) absorption and loss of reagents on the channel walls is prevented.
Once droplets of this kind have been generated and processed, it is necessary to extract the required analytical information. In this respect the detection method of choice should be rapid, provide high-sensitivity and low limits of detection, be applicable to a range of molecular species, be non-destructive and be able to be integrated with microfluidic devices in a facile manner. To address this need we have developed a suite of experimental tools and protocols that enable the extraction of large amounts of photophysical information from small-volume environments, and are applicable to the analysis of a wide range of physical, chemical and biological parameters. Herein two examples of these methods are presented and applied to the detection of single cells and the mapping of mixing processes inside picoliter-volume droplets. We report the entire experimental process including microfluidic chip fabrication, the optical setup and the process of droplet generation and detection.

Droplet-based microfluidic platforms are promising candidates for high throughput experimentation since they are able to generate picoliter, self-compartmentalized vessels inexpensively at kHz rates. Through integration with fast, sensitive and high resolution fluorescence spectroscopic methods, the large amounts of information generated within these systems can be efficiently extracted, harnessed and utilized.

Mikroakışkan damlacık platformları için Floresan algılama yöntemleri

The development of microfluidic platforms for performing chemistry and biology has in large part been driven by a range of potential benefits that ...

Biyomühendislik

Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets

Here, we demonstrate a simple method for the rapid production of size-controllable, monodisperse, W/O microdroplets using a capillary-based centrifugal microfluidic device. W/O microdroplets have recently been used in powerful methods that enable miniaturized chemical experiments. Therefore, developing a versatile method to yield monodisperse W/O microdroplets is needed. We have developed a method for generating monodisperse W/O microdroplets based on a capillary-based centrifugal axisymmetric co-flowing microfluidic device. We succeeded in controlling the size of microdroplets by adjusting the capillary orifice. Our method requires equipment that is easier-to-use than with other microfluidic techniques, requires only a small volume (0.1-1 &#181;l) of sample solution for encapsulation, and enables the production of hundreds of thousands number of W/O microdroplets per second. We expect this method will assist biological studies that require precious biological samples by conserving the volume of the samples for rapid quantitative analysis biochemical and biological studies.

Here, we demonstrate a simple production method for size-controllable, monodisperse, water-in-oil (W/O) microdroplets using a capillary-based centrifugal microfluidic device. This method requires only a small sample volume and enables high-yield production. We expect this method will be useful for rapid biochemical and cellular analyses.

Tek dağılımlı mikro damlacıklar boyutu kontrollü Formasyonu için Kılcal tabanlı Santrifüj mikroakışkan Cihaz

Here, we demonstrate a simple method for the rapid production of size-controllable, monodisperse, W/O microdroplets using a capillary-based centrifugal ...

Mühendislik

Double Emulsion Generation Using a Polydimethylsiloxane (PDMS) Co-axial Flow Focus Device

Double emulsions are useful in a number of biological and industrial applications in which it is important to have an aqueous carrier fluid. This paper presents a polydimethylsiloxane (PDMS) microfluidic device capable of generating water/oil/water double emulsions using a coaxial flow focusing geometry that can be fabricated entirely using soft lithography. Similar to emulsion devices using glass capillaries, double emulsions can be formed in channels with uniform wettability and with dimensions much smaller than the channel sizes. Three dimensional flow focusing geometry is achieved by casting a pair of PDMS slabs using two layer soft lithography, then mating the slabs together in a clamshell configuration. Complementary locking features molded into the PDMS slabs enable the accurate registration of features on each of the slab surfaces. Device testing demonstrates formation of double emulsions from 14 &#181;m to 50 &#181;m in diameter while using large channels that are robust against fouling and clogging.

Double Emulsion Generation Using a Polydimethylsiloxane PDMS Co-axial Flow Focus Device

Microfluidic double emulsions generation typically involves devices with patterned wettability or custom-fabricated glass components. Here we describe the fabrication and testing of an all polydimethylsiloxane (PDMS) double emulsion generator that does not require surface treatment or complicated fabrication processes, and is capable of producing double emulsions down to 14 &#181;m.

Bir Polidimetilsiloksan (PDMS) Koaksiyel Akış Odak Aygıtı Kullanma Çift Emülsiyon Üretimi

Double emulsions are useful in a number of biological and industrial applications in which it is important to have an aqueous carrier fluid. This paper ...

A Femtoliter Droplet Array for Massively Parallel Protein Synthesis from Single DNA Molecules

Advances in spatial resolution and detection sensitivity of scientific instrumentation make it possible to apply small reactors for biological and chemical research. To meet the demand for high-performance microreactors, we developed a femtoliter droplet array (FemDA) device and exemplified its application in massively parallel cell-free protein synthesis (CFPS) reactions. Over one million uniform droplets were readily generated within a finger-sized area using a two-step oil-sealing protocol. Every droplet was anchored in a femtoliter microchamber composed of a hydrophilic bottom and a hydrophobic sidewall. The hybrid hydrophilic-in-hydrophobic structure and the dedicated sealing oils and surfactants are crucial for stably retaining the femtoliter aqueous solution in the femtoliter space without evaporation loss. The femtoliter configuration and the simple structure of the FemDA device allowed minimal reagent consumption. The uniform dimension of the droplet reactors made large-scale quantitative and time-course measurements convincing and reliable. The FemDA technology correlated the protein yield of the CFPS reaction with the number of DNA molecules in each droplet. We streamlined the procedures about the microfabrication of the device, the formation of the femtoliter droplets, and the acquisition and analysis of the microscopic image data. The detailed protocol with the optimized low running cost makes the FemDA technology accessible to everyone who has standard cleanroom facilities and a conventional fluorescence microscope in their own place.

The overall goal of the protocol is to prepare over one million ordered, uniform, stable, and biocompatible femtoliter droplets on a 1 cm2 planar substrate that can be used for cell-free protein synthesis.

Tek DNA Moleküllerinden Kitlesel Paralel Protein Sentezi için Femtolitre Damlacık Dizisi

Advances in spatial resolution and detection sensitivity of scientific instrumentation make it possible to apply small reactors for biological and ...

Kimya

Fabricating Degradable Thermoresponsive Hydrogels on Multiple Length Scales via Reactive Extrusion, Microfluidics, Self-assembly, and Electrospinning

While various smart materials have been explored for a variety of biomedical applications (e.g., drug delivery, tissue engineering, bioimaging, etc.), their ultimate clinical use has been hampered by the lack of biologically-relevant degradation observed for most smart materials. This is particularly true for temperature-responsive hydrogels, which are almost uniformly based on polymers that are functionally non-degradable (e.g., poly(N-isopropylacrylamide) (PNIPAM) or poly(oligoethylene glycol methacrylate) (POEGMA)). As such, to effectively translate the potential of thermoresponsive hydrogels to the challenges of remote-controlled or metabolism-regulated drug delivery, cell scaffolds with tunable cell-material interactions, theranostic materials with the potential for both imaging and drug delivery, and other such applications, a method is required to render the hydrogels (if not fully degradable) at least capable of renal clearance following the required lifetime of the material. To that end, this protocol describes the preparation of hydrolytically-degradable hydrazone-crosslinked hydrogels on multiple length scales based on the reaction between hydrazide and aldehyde-functionalized PNIPAM or POEGMA oligomers with molecular weights below the renal filtration limit. Specifically, methods to fabricate degradable thermoresponsive bulk hydrogels (using a double barrel syringe technique), hydrogel particles (on both the microscale through the use of a microfluidics platform facilitating simultaneous mixing and emulsification of the precursor polymers and the nanoscale through the use of a thermally-driven self-assembly and cross-linking method), and hydrogel nanofibers (using a reactive electrospinning strategy) are described. In each case, hydrogels with temperature-responsive properties similar to those achieved via conventional free radical cross-linking processes can be achieved, but the hydrazone cross-linked network can be degraded over time to re-form the oligomeric precursor polymers and enable clearance. As such, we anticipate these methods (which may be generically applied to any synthetic water-soluble polymer, not just smart materials) will enable easier translation of synthetic smart materials to clinical applications.

Protocols are described for the fabrication of degradable thermoresponsive hydrogels based on hydrazone cross-linking of polymeric oligomers on the bulk scale, microscale, and nanoscale, the latter for preparation of both gel nanoparticles and nanofibers.

Parçalanabilir Thermoresponsive Hydrogels birden fazla uzunluğu ölçeklerde yolu ile reaktif ekstrüzyon, havacilik, kendinden montajlı imalatı ve Electrospinning

While various smart materials have been explored for a variety of biomedical applications (e.g., drug delivery, tissue engineering, bioimaging, etc.), ...

Image-guided, Laser-based Fabrication of Vascular-derived Microfluidic Networks

This detailed protocol outlines the implementation of image-guided, laser-based hydrogel degradation for the fabrication of vascular-derived microfluidic networks embedded in PEGDA hydrogels. Here, we describe the creation of virtual masks that allow for image-guided laser control; the photopolymerization of a micromolded PEGDA hydrogel, suitable for microfluidic network fabrication and pressure head-driven flow; the setup and use of a commercially available laser scanning confocal microscope paired with a femtosecond pulsed Ti:S laser to induce hydrogel degradation; and the imaging of fabricated microfluidic networks using fluorescent species and confocal microscopy. Much of the protocol is focused on the proper setup and implementation of the microscope software and microscope macro, as these are crucial steps in using a commercial microscope for microfluidic fabrication purposes that contain a number of intricacies. The image-guided component of this technique allows for the implementation of 3D image stacks or user-generated 3D models, thereby allowing for creative microfluidic design and for the fabrication of complex microfluidic systems of virtually any configuration. With an expected impact in tissue engineering, the methods outlined in this protocol could aid in the fabrication of advanced biomimetic microtissue constructs for organ- and human-on-a-chip devices. By mimicking the complex architecture, tortuosity, size, and density of in vivo vasculature, essential biological transport processes can be replicated in these constructs, leading to more accurate in vitro modeling of drug pharmacokinetics and disease.

This protocol outlines the implementation of image-guided, laser-based hydrogel degradation to fabricate vascular-derived, biomimetic microfluidic networks embedded in poly(ethylene glycol) diacrylate (PEGDA) hydrogels. These biomimetic microfluidic systems may be useful for tissue engineering applications, generation of in vitro disease models, and fabrication of advanced "on-a-chip" devices.

Vasküler kaynaklı mikroakışkan Ağların görüntü kılavuzluğunda, Lazer tabanlı Fabrikasyon

This detailed protocol outlines the implementation of image-guided, laser-based hydrogel degradation for the fabrication of vascular-derived microfluidic ...

Glass-Based Devices to Generate Drops and Emulsions

In this manuscript, three different step-by-step protocols to generate highly monodisperse emulsion drops using glass-based microfluidics are described. The first device is built for the generation of simple drops driven by gravity. The second device is designed to generate emulsion drops in a coflowing scheme. The third device is an extension of the coflowing device with the addition of a third liquid that acts as an electric ground, allowing the formation of electrified drops that subsequently discharge. In this setup, two of the three liquids have an appreciable electrical conductivity. The third liquid mediates between these two and is a dielectric. A&#160;voltage difference applied between the two conducting liquids creates an electric field that couples with hydrodynamic stresses of the coflowing liquids, affecting the jet and drop formation process. The addition of the electric field provides a path to generate smaller drops than in simple coflow devices and for generating particles and fibers with a wide range of sizes.

Here, a protocol to manufacture glass-based microfluidic devices used for generating highly monodisperse emulsions with controlled drop size is presented.

Damla ve Emülsiyon Üretmek için Cam Bazlı Cihazlar

In this manuscript, three different step-by-step protocols to generate highly monodisperse emulsion drops using glass-based microfluidics are described. ...

Multi-step Variable Height Photolithography for Valved Multilayer Microfluidic Devices

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

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

Ventilli Katmanlı mikroakışkan Cihazlar için çok adımlı Değişken Yükseklik Fotolitografi

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

Generation of Dynamical Environmental Conditions using a High-Throughput Microfluidic Device

Mimicking in vivo environmental conditions is crucial for in vitro studies on complex life machinery. However, current techniques targeting live cells and organs are either highly expensive, like robotics, or lack nanoliter volume and millisecond time accuracy in liquid manipulation. We herein present the design and fabrication of a microfluidic system, which consists of 1,500 culture units, an array of enhanced peristaltic pumps and an on-site mixing modulus. To demonstrate the capacities of the microfluidic device, neural stem cell (NSC) spheres are maintained in the proposed system. We observed that when the NSC sphere is exposed to CXCL in day 1 and EGF in day 2, the round-shaped conformation is well maintained. Variation in the input order of 6 drugs causes morphological changes to the NSC sphere and the expression level representative marker for NSC stemness (i.e., Hes5 and Dcx). These results indicate that dynamic and complex environmental conditions have great effects on NSC differentiation and self-renewal, and the proposed microfluidic device is a suitable platform for high throughput studies on the complex life machinery.

We present a microfluidic system for high throughput studies on complex life machinery, which consists of 1500 culture units, an array of enhanced peristaltic pumps and an on-site mixing modulus. The microfluidic chip allows for the analysis of the highly complex and dynamic micro-environmental conditions in vivo.

Yüksek Verimli Mikroakışkan Cihaz Kullanarak Dinamik ÇevreSel Koşulların Üretilmesi

Mimicking in vivo environmental conditions is crucial for in vitro studies on complex life machinery. However, current techniques targeting live cells and ...

Kombinatoryal Plug Üretimi için Mikroakışkan Bir Cihazın Kurulumu

Bilayer Microfluidic Device for Combinatorial Plug Production

Droplet microfluidics is a versatile tool that allows the execution of a large number of reactions in chemically distinct nanoliter compartments. Such systems have been used to encapsulate a variety of biochemical reactions - from incubation of single cells to implementation of PCR reactions, from genomics to chemical synthesis. Coupling the microfluidic channels with regulatory valves allows control over their opening and closing, thereby enabling the rapid production of large-scale combinatorial libraries consisting of a population of droplets with unique compositions. In this paper, protocols for the fabrication and operation of a pressure-driven, PDMS-based bilayer microfluidic device that can be utilized to generate combinatorial libraries of water-in-oil emulsions called plugs are presented. By incorporating software programs and microfluidic hardware, the flow of desired fluids in the device can be controlled and manipulated to generate combinatorial plug libraries and to control the composition and quantity of constituent plug populations. These protocols will expedite the process of generating combinatorial screens, particularly to study drug response in cells from cancer patient biopsies.

Yazar Spotlight: Hesaplamalı ve Deneysel Yaklaşımları Hassas Onkolojiye Entegre Etme

The fabrication of a polydimethylsiloxane (PDMS)-based bilayer device for the production of combinatorial libraries in water-in-oil emulsions (plugs) is presented here. The necessary hardware and software required to automate plug production are detailed in the protocol, and the production of a quantitative library of fluorescent plugs is also demonstrated.

Kombinatoryal Fiş Üretimi için İki Katmanlı Mikroakışkan Cihaz

Droplet microfluidics is a versatile tool that allows the execution of a large number of reactions in chemically distinct nanoliter compartments. Such ...