وقد تم دعم هذا العمل من قبل كلية أوغدن للعلوم والهندسة، بما في ذلك الدعم الداخلي من قسم الكيمياء ومن مكتب البحوث (اللجنة الملكية 13-8032) في جامعة كنتاكي الغربية. وكانت المساعدة من الدكتور جون Andersland في مرفق WKU المجهري (صور SEM) وأستاذ مشارك يان تساو معهد WKU للاحتراق علوم والهندسة (تحليل BET) المركزي لإجراء هذا العمل.

1. إعداد مستحلب <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> مستحلب المحتويات <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> كتلة كمية مناسبة من الملح لإنتاج 10 مل من 0.03-M حل. لالبلاتين (IV) قياس كلوريد 0.101 غرام، لالزنك (II) كلوريد (ZnCl 2) قياس 0.032 غرام، وكلوريد الصوديوم (كلوريد الصوديوم) قياس 0،018 غرام. </li><li style=";text-align:right;direction:rtl"> في أنابيب الاختبار الفردية، ويحل كل الملح في 10 مل من الماء DI. توضع جانبا لاستخدامها لاحقا. </li><li style=";text-align:right;direction:rtl"> استخدام 20 مل، قابل للغلق قارورة زجاجية عن محتويات طوال هذا الإجراء. الفارغة على التوازن في قارورة زجاجية. </li><li style=";text-align:right;direction:rtl"> تزن منتهية الفينيل polydimethylsiloxane بصب ببطء على قضيب ضجة وداخل قارورة زجاجية يستريح على نطاق وركزت. وزن من 1.02 غرام (أي ما يعادل 1.080 مل). ملاحظة: اللزوجة العالية لهذا البوليمر يجعل pipetting لعملي. </li><li style=";text-align:right;direction:rtl"> ماصة 1.02 مل من ن الهيبتان إلى القارورة. إضافة 2 قطرات من السطحي غير الأيونية (صوربيتان monoleate) إلى القارورة. ماصة 0.3 مل من محلول الملح و 0.45 مل من DI-الماء إلى قارورة. </li><li style=";text-align:right;direction:rtl"> ختم قارورة زجاجية قبل الشد الغطاء بإحكام. هزة بقوة لمدة 60 ثانية لبدء مستحلب قبل البدء صوتنة. </li></ol></li><li style=";text-align:right;direction:rtl"> بناء المياه حمام جهاز Sonicator <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ملء sonicator مع الماء تصل إلى الحد الأدنى من خط التعبئة. إضافة 250 مل من ماء الصنبور إلى كوب 400 مل. ملء كوب 400 مل مع الثلج بحيث أن مستوى الماء فقط عند الحافة. </li><li style=";text-align:right;direction:rtl"> ضع هذه الكأس داخل المياه sonicator حمام. تحقق من خط التعبئة، على sonicator، وتعديل إذا لزم الأمر. وضع عصابة تقف مباشرة بجوار sonicator المياه حمام. </li><li style=";text-align:right;direction:rtl"> باستخدام اثنين من عصابة تقف والمشابك، والموقف منها بحيث يتم تمديد ذراع خارج، عمودي على الحلبة الوقوف وتمديد واحدة أخرى نحو حمام مائي بحيث يشير إلى أسفل في كوب مملوء بالماء الجليد. نعلق المشبك أخرى إلى الحلبة الوقوف مع ميزان حرارة أسفل في كوب 400 مل بحيث يمكن رصد درجة الحرارة في جميع أنحاء صوتنة. </li></ol></li><li style=";text-align:right;direction:rtl"> Emulsifالإجراءات ication <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ضع القارورة التي تحتوي على مستحلب بحيث يتم مغمورة بالكامل في كوب 400 مل من خلال تأمين عليه في المشبك جاحظ أسفل إلى الدورق. </li><li style=";text-align:right;direction:rtl"> تأكد من أن قارورة زجاجية تحتوي على خليط PDMS لا لمس جانبي الكأس للقضاء على الحرارة الناتجة عن الاحتكاك. </li><li style=";text-align:right;direction:rtl"> بدوره على sonicator وضبط الوقت صوتنة لمدة 7 دقائق. لأن مستحلب هو الحرارة جدا حساسة، تأكد من أن درجة الحرارة داخل الدورق ما بين 0 و 5 درجات مئوية طوال صوتنة. بدء صوتنة بمجرد أن درجة الحرارة داخل الدورق أمر مرغوب فيه. </li><li style=";text-align:right;direction:rtl"> بعد 7 دقائق من صوتنة، إزالة القارورة وبلطف يهز / دوامة لمدة 1 دقيقة، وعقد قمة القارورة للقضاء على أي كتل التي يمكن أن تشكل في مستحلب. </li><li style=";text-align:right;direction:rtl"> تخلص من محتويات الدورق. إعادة ملء مع 250 مل من الماء وإضافة الجليد إلى التعبئة لفي حدود 1 سم أعلى الدورق. </li><li style=";text-align:right;direction:rtl"> ضع القارورة مرة أخرى في المشبك. يغرق تحت الماء المثلج لresumالبريد صوتنة لمدة 7 دقائق. </li><li style=";text-align:right;direction:rtl"> كرر الخطوات من 1.3.3 إلى 1.3.6 ليصبح المجموع فترات صوتنة ثمانية، 7 دقيقة، أو حتى صوتنة يبدو أن متجانسة وعدم وجود كتل موجودة. تخزين في RT. ملاحظة: يجب أن يكون استحلاب مستقرة لعدة أيام، ولكن يمكن استردادها من قبل صوتنة على النحو الوارد أعلاه في 7 دقيقة فترات حتى يبدو متجانسا. تخزين في RT. </li></ol></li></ol> 2. عبر ربط <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> الإعداد لإضافة Triethoxysilane / السطحي الحل <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ماصة 5.4 مل triethoxysilane في أنبوب الاختبار. ضع في أنبوب اختبار رف تحت غطاء محرك السيارة لاستخدامها لاحقا. </li><li style=";text-align:right;direction:rtl"> ملء كوب 400 مل مع الماء المثلج ووضعه تحت غطاء محرك السيارة. التالي إلى المكان كوب 400 مل حلقة تقف مع المشبك المرفقة، بمد مباشرة عبر افتتاح الكأس. وسيكون هذا هو حمام كريم لإضافة triethoxysilane. </li><li style=";text-align:right;direction:rtl"> وضع صفيحة ساخنة على الجانب الآخر من موقف حلقة. ملء كوب 800 مل مع ما يقرب من 700 مل منماء الصنبور. وضعه على طبق ساخن. </li><li style=";text-align:right;direction:rtl"> بدوره على طبق ساخن والحفاظ على درجة حرارة 75 إلى 85 درجة مئوية داخل كوب 800 مل. إرفاق المشبك مع ميزان حرارة لموقف حلقة حتى درجة حرارة المياه داخل الدورق 800 مل يمكن رصدها. </li><li style=";text-align:right;direction:rtl"> إنتاج حل للتوتر السطحي عن طريق إذابة 0.5 غرام من دوديسيل كبريتات الصوديوم إلى 375 مل من الماء (4.62 ملم). إضافة حوالي 10 مل من محلول السطحي لنظيف، فارغة أنبوب الاختبار. </li><li style=";text-align:right;direction:rtl"> نعلق المشبك أخرى إلى الحلبة الوقوف تحت غطاء محرك السيارة مع أنبوب الاختبار السطحي المضمون بحيث مستوياته السائل تحت سطح الماء داخل الدورق 800 مل. سماح 10 دقيقة للموازنة الحرارية. </li><li style=";text-align:right;direction:rtl"> الرطب قطعة من ورق الترشيح ووضعه في الجزء العلوي من قمع صغير. ضع الجذعية من القمع داخل دورق مخروطي 250 مل ووضع قارورة تحت غطاء محرك السيارة. </li></ol></li><li style=";text-align:right;direction:rtl"> إضافة Triethoxysilane <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ضع قارورة مستحلب في المشبك على منقار مياه الجليدإيه داخل غطاء محرك السيارة. ضع الخليط PDMS في المشبك بحيث محتويات قنينة هي تحت سطح الماء. إضافة triethoxysilane تسبب رد فعل الطاردة للحرارة، لذلك يجب أن تبقى مستحلب الباردة من أجل الحفاظ على هيكلها. </li><li style=";text-align:right;direction:rtl"> إزالة الغطاء من قارورة زجاجية لتجنب الغازي بناء. </li><li style=";text-align:right;direction:rtl"> صب ببطء أنبوب اختبار يحتوي على triethoxysilane في قارورة زجاجية في تيار مستمر على مدى ما يقرب من 10 ثانية (حوالي 0.5 مل / ثانية). تنبيه: إضافة triethoxysilane ء تفاعل طارد للحرارة والإفراج عن كلوريد الهيدروجين الكاوية (حمض الهيدروكلوريك) الغاز. سوف القارورة تصبح ساخنة للغاية وسوف الغازات السامة تتطور من الخليط. لا يحرك محتويات أثناء إضافة triethoxysilane. </li><li style=";text-align:right;direction:rtl"> بعد إضافة triethoxysilane تماما، وإثارة بلطف محتويات مع ضجة قضيب الزجاج بينما يرتدي القفازات الواقية الحرارة. الانتظار لمدة 2 دقيقة أو حتى توقف الغاز المتطورة من القارورة. </li><li style=";text-align:right;direction:rtl"> الفلبسبب عبر ربط، لا توجد مرحلة فصل واضح في العينة. إذا كتل موجودة، وختم قارورة ويهز صارم لمدة 20 ثانية في حين عقد القارورة التي كتبها الغطاء. </li></ol></li><li style=";text-align:right;direction:rtl"> إنتاج حبة <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> استخدام نظيفة، الزجاج ماصة باستير لرسم مستحلب عبر ربط من قارورة زجاجية. إضافة قطرة من الحكمة في حل السطحي (الذي يجب أن يكون قائما بين 75 و 85 درجة مئوية) في أنبوب الاختبار يأخذ أقل وقت ممكن في ما بين قطرات مستحلب عبر ربط. </li><li style=";text-align:right;direction:rtl"> 30 ثانية إلى 1 دقيقة بعد إضافة مستحلب، فإن الحل السطحي تبدأ ببطء لتطوير الغاز حيث تبدأ المواد الصلبة لتشكيل داخل أنبوب الاختبار. </li><li style=";text-align:right;direction:rtl"> في حين يرتدي قفازات واقية للحرارة، واتخاذ أنبوب الاختبار من المشبك وتصب محتوياته بالكامل في جهاز الترشيح تحت غطاء محرك السيارة. مرشح لمدة 5 دقائق. إزالة ورقة الترشيح من مرشح. </li><li style=";text-align:right;direction:rtl"> نقل المواد الصلبة التي تمت تصفيتها على كوب ساعة وحبات منفصلة للO / N التجفيف تحت حالعود. يمكن تأجيل تنظيف حبات أجل غير مسمى. متجر الخرز في RT تجفف في قارورة زجاجية محكمة الغلق لحين الحاجة إليها، ونظيفة مباشرة قبل استخدامها. </li></ol></li><li style=";text-align:right;direction:rtl"> حبة التنظيف <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> إنشاء جهاز الترشيح آخر تحت غطاء محرك السيارة ووضع حبات المجففة داخل القمع على رأس ورقة الترشيح. </li><li style=";text-align:right;direction:rtl"> استخدام زجاجة غسل بلاستيكية مملوءة DI-الماء لشطف حبات بلطف، وتتحرك حول لهم قليلا لضمان تشطف كل الخرز. </li><li style=";text-align:right;direction:rtl"> السماح حبات جافة لمدة 1 ساعة عن طريق وضعها على الزجاج ووتش تحت غطاء محرك السيارة. استخدام زجاجة غسل مليئة hexanes لشطف الخرز باستخدام نفس الأسلوب لالشطف بالماء. </li><li style=";text-align:right;direction:rtl"> وضع الخرز على الزجاج ووتش. وضع الزجاج ووتش والخرز تحت غطاء محرك السيارة لتجف. </li><li style=";text-align:right;direction:rtl"> بعد حبات جافة تماما، ووضعها في قارورة زجاجية صغيرة قابل للغلق وتخزينها في RT لاستخدامها في المستقبل. </li></ol></li><li style=";text-align:right;direction:rtl"> تركيب الخرز للإعدادات تحليل SEM وSEM <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> وضع شريط من تأليفالكربون شريط موصل uble من جانب وعلى رأس كعب على الذي يقام الخرز. باستخدام مقص، وتقليم حول كعب لضمان عدم توقف الشريط على الحواف. </li><li style=";text-align:right;direction:rtl"> ضع قطعة من ورق الترشيح تحت كعب على سطح مستو. إزالة الطبقة العليا من الشريط بحيث يتعرض الجانب السفلي لاصقة. </li><li style=";text-align:right;direction:rtl"> صب بلطف الخرز على كعب. وبعض حبات العصا إلى الشريط، ولكن معظم سوف ترتد والأرض على ورقة الترشيح. صب هذه العودة إلى القارورة حتى لو بقيت على ورقة الترشيح. كرر إذا لزم الأمر، وغسل أي الخرز (وفقا ل2.4.2 إلى 2.4.5) التي تصبح ملوثة. </li><li style=";text-align:right;direction:rtl"> لضمان حبات آمنة على كعب، استخدم حقنة لمبة وضربة طفيفة عن كثب على سطح كعب. صب مزيد من الخرز على كعب إلا إذا كان عدد قليل انضمت إلى الشريط. التأكد من أن جميع حبات آمنة قبل وضع كعب في غرفة SEM واخلاء عليه. </li><li style=";text-align:right;direction:rtl"> مرة واحدة وقد شنت العينات بشكل صحيح أنهم الآن على استعداد للخضوع SEM الشرجيسيس 15. جمع الصور في وضع LOW VAC في 15 كيلو لتحسين القرار من الميزات سطح حبة ل. </li></ol></li></ol>

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J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Concave+porosity+non-polar+beads+by+a+modified+microbubble+fabrication.">Concave porosity non-polar beads by a modified microbubble fabrication.</a> Mater. Lett. 98, 105-107 (2013).</li><li>Flegler, S. L., Heckman, J. W., Klomparens, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Scanning and Transmission Electron Microscopy: An Introduction. , 151-155 (1995).</li><li>Brunauer, S., Emmett, P. H., Teller, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adsorption+of+Gases+in+Multimolecular+Layers.">Adsorption of Gases in Multimolecular Layers.</a> J. Am. Chem. Soc. 60 (2), 309-319 (1938).</li><li>Sing, K. S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+porous+materials:+past,+present+and+future.">Characterization of porous materials: past, present and future.</a> Colloids Surf. A. 241 (1), 3-7 (2004).</li></ol>

The authors have nothing to disclose.

حبات المنتجة باستخدام هذا البروتوكول (وعن طريق ضبط تركيز بالكهرباء والهوية) تختلف جوهريا عن تلك المنتجة مع قوة مستحلب المنخفض الأيونية، كما رأينا من خلال المقارنة بين الشكل 1A إلى الصور SEM أخرى في الشكل 1. لدينا التقرير الأولي تستخدم شركة ال...

Polydimethylsiloxane (PDMS) is one of the most widely used silicone compounds. Its biocompatibility has led to widespread use in implant and other biomedical engineering structures1,2. It is trivially cross-linked into elastic structures using an organosilyl compound (such as triethoxysilane), a simple and reliable procedure which has made it useful for cast polymer applications where some flexibility is required3. Once cross-linked, PDMS is largely inert, particularly in biological conditions, and is therefore useful for a variety of food and medical applications4,5. Ease of casting, chemical inertness, and hydrophobicity have made it a natural choice for microfluidic devices6,7. Its affinity for non-halogenated, non-polar organic compounds has made it a popular stationary phase in separations chemistry8-10. 
Recently, microbubble fabrications have been used to generate porous beads for use as catalyst structural substrates or in chemical separations11,12. In both applications, ideal materials will have a maximized surface-area-to-volume (SAV) ratio for best efficiency. In a microbubble fabrication process, microstructuring of materials is typically accomplished by isolating the polymer in aliphatic &#8220;microbubbles&#8221; by emulsification in an aqueous continuous phase. The initial report of microporous PDMS beads produced them by mechanical emulsification of two phases (aliphatic and aqueous)13. The stock PDMS liquid (and its cross-linking agent) is dissolved into the aliphatic phase, which is structured into microscopic beads by being forced to cavitate within the (continuous) aqueous phase. The emulsification is stabilized by the addition of a non-ionic surfactant. When the emulsion is added dropwise to a heated bath, solid beads form by agglomeration of the microbubbles into clusters of tiny spheres of cross-linked PDMS. Our goal in this protocol is to modify this procedure to develop beads with an inverted porosity to improve the SAV ratio of the material.
As reported previously, control of the beads can be directed to some extent by the aliphatic:aqueous ratios in the emulsion. However, we have reported recently that addition of platinum(IV) chloride (PtCl4) inverts the porosity: materials are formed in which the PDMS is riddled with concave pores14. This indicates that the aqueous layer cavitates inside the aliphatic one, despite having similar aliphatic:aqueous ratios to those published in the original work13. The primary advantage of our method is that this concave porosity should naturally result in an increased SAV ratio, and thus, improved efficiency for analytical chemistry applications. While we are continuing to explore the specific effects of the addition of the platinum compound, we show here that the same effect can be accomplished using any aqueously soluble ionic compound, though perhaps to a reduced extent. Because our techniques also differ in some key aspects from what has been previously reported, we present our protocols here as a video to encourage others to extend our methods. Most notably, we use a common bath sonicator of the type used to clean glassware or other equipment, rather than the (considerably more expensive) probe sonicator often used in microbubble fabrication. This adjusted approach to the microbubble fabrication procedure could potentially be extended for the production of large quantities of bulk materials as well, creating porous sheets or slabs which could have applications for biomedical devices, aerospace and automotive industry, or substrates for chemical catalysis. Users seeking to generate high-SAV-ratio, microstructured materials using other similar polymers for such analyses may find that our protocols can be extended to any polymer for which the microbubble emulsion technique can be applied.

<table><tbody><tr><td>Poly(dimethylsiloxane), vinyl terminated</td><td>Sigma-Aldrich</td><td>68083-19-2</td><td></td></tr><tr><td>n-Heptane</td><td>Sigma-Aldrich</td><td>142-82-5</td><td>Flammable</td></tr><tr><td>Triethoxysilane</td><td>Sigma-Aldrich</td><td>998-30-1</td><td>Flammable, Accutely Toxic</td></tr><tr><td>Sorbitan Monoleate (Span-80)</td><td>Fluker</td><td>1338-43-8</td><td></td></tr><tr><td>Platinum(IV) Chloride</td><td>Sigma-Aldrich</td><td>13454-96-1</td><td>Accutely Toxic</td></tr><tr><td>Zinc(II) Chloride</td><td>Sigma-Aldrich</td><td>7646-85-7</td><td></td></tr><tr><td>Sodium Chloride</td><td>Sigma-Aldrich</td><td>7647-14-5</td><td></td></tr><tr><td>2.8 L Water Bath Sonicator</td><td>VWR</td><td>97043-964</td><td></td></tr></tbody></table>

microbubble fabrication of concave-porosity pdms beads

Microbubble fabrication (by use of a fine emulsion) provides a means of increasing the surface-area-to-volume (SAV) ratio of polymer materials, which is particularly useful for separations applications. Porous polydimethylsiloxane (PDMS) beads can be produced by heat-curing such an emulsion, allowing the interface between the aqueous and aliphatic phases to mold the morphology of the polymer. In the procedures described here, both polymer and crosslinker (triethoxysilane) are sonicated together in a cold-bath sonicator. Following a period of cross-linking, emulsions are added dropwise to a hot surfactant solution, allowing the aqueous phase of the emulsion to separate, and forming porous polymer beads. We demonstrate that this method can be tuned, and the SAV ratio optimized, by adjusting the electrolyte content of the aqueous phase in the emulsion. Beads produced in this way are imaged with scanning electron microscopy, and representative SAV ratios are determined using Brunauer&#8211;Emmett&#8211;Teller (BET) analysis. Considerable variability with the electrolyte identity is observed, but the general trend is consistent: there is a maximum in SAV obtained at a specific concentration, after which porosity decreases markedly.

وتظهر الصور SEM تمثيلية من الخرز الناشئة من المستحلبات مع الظروف بالكهرباء مختلفة في الشكل 1. ويبين الشكل 1A حبة مشابهة لتلك التي حصلت عليها DuFaud، وآخرون. 13، المنتجة باستخدام إجراءاتنا، دون إضافة أي بالكهرباء. الخرز هو مبي?...

Watch this Scientific Journal Video about Microbubble Fabrication of Concave-porosity PDMS Beads at JoVE.com

Microbubble Fabrication of Concave-porosity PDMS Beads

Procedures used to generate microstructured concave-porosity polydimethylsiloxane beads are presented. Effects of electrolyte concentration and identity within the aqueous phase are particularly emphasized.

تصنيع Microbubble من الخرز PDMS مقعرة المسامية

Microbubble fabrication (by use of a fine emulsion) provides a means of increasing the surface-area-to-volume (SAV) ratio of polymer materials, which is ...

microbubble-fabrication-of-concave-porosity-pdms-beads

Research

JoVE Journal

Chemistry

8.3K Views.  Western Kentucky University. Procedures used to generate microstructured concave-porosity polydimethylsiloxane beads are presented. Effects of electrolyte concentration and identity within the aqueous phase are particularly emphasized. 

Video: Microbubble Fabrication of Concave-porosity PDMS Beads

Microfluidic Chips Controlled with Elastomeric Microvalve Arrays

Miniaturized microfluidic systems provide simple and effective solutions for low-cost point-of-care diagnostics and high-throughput biomedical assays. Robust flow control and precise fluidic volumes are two critical requirements for these applications. We have developed microfluidic chips featuring elastomeric polydimethylsiloxane (PDMS) microvalve arrays that: 1) need no extra energy source to close the fluidic path, hence the loaded device is highly portable; and 2) allow for microfabricating deep (up to 1 mm) channels with vertical sidewalls and resulting in very precise features.

The PDMS microvalves-based devices consist of three layers: a fluidic layer containing fluidic paths and microchambers of various sizes, a control layer containing the microchannels necessary to actuate the fluidic path with microvalves, and a middle thin PDMS membrane that is bound to the control layer. Fluidic layer and control layers are made by replica molding of PDMS from SU-8 photoresist masters, and the thin PDMS membrane is made by spinning PDMS at specified heights. The control layer is bonded to the thin PDMS membrane after oxygen activation of both, and then assembled with the fluidic layer. The microvalves are closed at rest and can be opened by applying negative pressure (e.g., house vacuum). Microvalve closure and opening are automated via solenoid valves controlled by computer software.

Here, we demonstrate two microvalve-based microfluidic chips for two different applications. The first chip allows for storing and mixing precise sub-nanoliter volumes of aqueous solutions at various mixing ratios. The second chip allows for computer-controlled perfusion of microfluidic cell cultures.

The devices are easy to fabricate and simple to control. Due to the biocompatibility of PDMS, these microchips could have broad applications in miniaturized diagnostic assays as well as basic cell biology studies.

We demonstrate protocols for manufacturing and automating elastomeric polydimethylsiloxane (PDMS)-based microvalve arrays that need no extra energy to close and feature photolithographically defined precise volumes. A parallel subnanoliter-volume mixer and an integrated microfluidic perfusion system are presented.

رقائق ميكروفلويديك مع الشواهد صالحة Microvalve المرنة

Miniaturized microfluidic systems provide simple and effective solutions for low-cost point-of-care diagnostics and high-throughput biomedical assays.  ...

Biology

Creating Sub-50 Nm Nanofluidic Junctions in PDMS Microfluidic Chip via Self-Assembly Process of Colloidal Particles

Polydimethylsiloxane (PDMS) is the prevailing building material to make microfluidic devices due to its ease of molding and bonding as well as its transparency. Due to the softness of the PDMS material, however, it is challenging to use PDMS for building nanochannels. The channels tend to collapse easily during plasma bonding. In this paper, we present an evaporation-driven self-assembly method of silica colloidal nanoparticles to create nanofluidic junctions with sub-50 nm pores between two microchannels. The pore size as well as the surface charge of the nanofluidic junction is tunable simply by changing the colloidal silica bead size and surface functionalization outside of the assembled microfluidic device in a vial before the self-assembly process. Using the self-assembly of nanoparticles with a bead size of 300 nm, 500 nm, and 900 nm, it was possible to fabricate a porous membrane with a pore size of ~45 nm, ~75 nm and ~135 nm, respectively. Under electrical potential, this nanoporous membrane initiated ion concentration polarization (ICP) acting as a cation-selective membrane to concentrate DNA by ~1,700 times within 15 min. This non-lithographic nanofabrication process opens up a new opportunity to build a tunable nanofluidic junction for the study of nanoscale transport processes of ions and molecules inside a PDMS microfluidic chip.

We propose a simple self-assembly technique of silica colloidal nanoparticles to create a nanofluidic junction between two microchannels in polydimethylsiloxane (PDMS). Using this technique, a nanoporous bead membrane with a pore size down to ~45 nm was built inside a microchannel and applied to electrokinetic preconcentration of DNA samples.

إنشاء 50 نيوتن متر الفرعية علم الموائع النانوي المفارق في PDMS ميكروفلويديك رقاقة عن طريق عملية التجميع الذاتي من الغروية الجسيمات

Polydimethylsiloxane (PDMS) is the prevailing building material to make microfluidic devices due to its ease of molding and bonding as well as its ...

Engineering

A Microfluidic Technique to Probe Cell Deformability

Here we detail the design, fabrication, and use of a microfluidic device to evaluate the deformability of a large number of individual cells in an efficient manner. Typically, data for ~102 cells can be acquired within a 1 hr experiment. An automated image analysis program enables efficient post-experiment analysis of image data, enabling processing to be complete within a few hours. Our device geometry is unique in that cells must deform through a series of micron-scale constrictions, thereby enabling the initial deformation and time-dependent relaxation of individual cells to be assayed. The applicability of this method to human promyelocytic leukemia (HL-60) cells is demonstrated. Driving cells to deform through micron-scale constrictions using pressure-driven flow, we observe that human promyelocytic (HL-60) cells momentarily occlude the first constriction for a median time of 9.3 msec before passaging more quickly through the subsequent constrictions with a median transit time of 4.0 msec per constriction. By contrast, all-trans retinoic acid-treated (neutrophil-type) HL-60 cells occlude the first constriction for only 4.3 msec before passaging through the subsequent constrictions with a median transit time of 3.3 msec. This method can provide insight into the viscoelastic nature of cells, and ultimately reveal the molecular origins of this behavior.

We demonstrate a microfluidics-based assay to measure the timescale for cells to transit through a sequence of micron-scale constrictions.

تقنية ميكروفلويديك للتحقيق التشوه خلية

Here we detail the design, fabrication, and use of a microfluidic device to evaluate the deformability of a large number of individual cells in an ...

Bioengineering

Combining Fluidic Devices with Microscopy and Flow Cytometry to Study Microbial Transport in Porous Media Across Spatial Scales

Understanding the transport, dispersion and deposition of microorganisms in porous media is a complex scientific task comprising topics as diverse as hydrodynamics, ecology and environmental engineering. Modeling bacterial transport in porous environments at different spatial scales is critical to better predict the consequences of bacterial transport, yet current models often fail to up-scale from laboratory to field conditions. Here, we introduce experimental tools to study bacterial transport in porous media at two spatial scales. The aim of these tools is to obtain macroscopic observables (such as breakthrough curves or deposition profiles) of bacteria injected into transparent porous matrices. At the small scale (10-1000 &#181;m), microfluidic devices are combined with optical video-microscopy and image processing to obtain breakthrough curves and, at the same time, to track individual bacterial cells at the pore scale. At larger scale, flow cytometry is combined with a self-made robotic dispenser to obtain breakthrough curves. We illustrate the utility of these tools to better understand how bacteria are transported in complex porous media such as the hyporheic zone of streams. As these tools provide simultaneous measurements across scales, they pave the way for mechanism-based models, critically important for upscaling. Application of these tools may not only contribute to the development of novel bioremediation applications but also shed new light on the ecological strategies of microorganisms colonizing porous substrates.

Breakthrough curves (BTCs) are efficient tools to study the transport of bacteria in porous media. Here we introduce tools based on fluidic devices in combination with microscopy and flow cytometric counting to obtain BTCs.

الجمع بين الأجهزة Fluidic مع المجهر والتدفق cytometry لدراسة النقل الميكروبي في وسائل الإعلام المسامية عبر المقاييس المكانية

Understanding the transport, dispersion and deposition of microorganisms in porous media is a complex scientific task comprising topics as diverse as ...

Environment

Fabrication of Refractive-index-matched Devices for Biomedical Microfluidics

The use of microfluidic devices has emerged as a defining tool for biomedical applications. When combined with modern microscopy techniques, these devices can be implemented as part of a robust platform capable of making simultaneous complementary measurements. The primary challenge created by the combination of these two techniques is the mismatch in refractive index between the materials traditionally used to make microfluidic devices and the aqueous solutions typically used in biomedicine. This mismatch can create optical artifacts near the channel or device edges. One solution is to reduce the refractive index of the material used to fabricate the device by using a fluorinated polymer such as MY133-V2000 whose refractive index is similar to that of water (n = 1.33). Here, the construction of a microfluidic device made out of MY133-V2000 using soft lithography techniques is demonstrated, using O2 plasma in conjunction with an acrylic holder to increase the adhesion between the MY133-V2000 fabricated device and the polydimethylsiloxane (PDMS) substrate. The device is then tested by incubating it filled with cell culture media for 24 h to demonstrate the ability of the device to maintain cell culture conditions during the course of a typical imaging experiment. Finally, quantitative phase microscopy (QPM) is used to measure the distribution of mass within the live adherent cells in the microchannel. This way, the increased precision, enabled by fabricating the device from a low index of refraction polymer such as MY133-V2000 in lieu of traditional soft lithography materials such as PDMS, is demonstrated. Overall, this approach for fabricating microfluidic devices can be readily integrated into existing soft lithography workflows in order to reduce optical artifacts and increase measurement precision.

This protocol describes the fabrication of microfluidic devices from MY133-V2000 to eliminate artifacts that often arise in microchannels due to the mismatching refractive indices between microchannel structures and an aqueous solution. This protocol uses an acrylic holder to compress the encapsulated device, improving adhesion both chemically and mechanically.

تصنيع الأجهزة المتطابقة من مؤشر الانكسار للطب الحيوي ميكروفلويديكس

The use of microfluidic devices has emerged as a defining tool for biomedical applications. When combined with modern microscopy techniques, these devices ...

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.

ضعف مستحلب الجيل باستخدام Polydimethylsiloxane (PDMS) المشارك المحوري تدفق التركيز الأجهزة

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 Microfluidic Platform for High-throughput Single-cell Isolation and Culture

Studying the heterogeneity of single cells is crucial for many biological questions, but is technically difficult. Thus, there is a need for a simple, yet high-throughput, method to perform single-cell culture experiments. Here, we report a microfluidic chip-based strategy for high-efficiency single-cell isolation (~77%) and demonstrate its capability of performing long-term single-cell culture (up to 7 d) and cellular heterogeneity analysis using clonogenic assay. These applications were demonstrated with KT98 mouse neural stem cells, and A549 and MDA-MB-435 human cancer cells. High single-cell isolation efficiency and long-term culture capability are achieved by using different sizes of microwells on the top and bottom of the microfluidic channel. The small microwell array is designed for precisely isolating single-cells, and the large microwell array is used for single-cell clonal culture in the microfluidic chip. This microfluidic platform constitutes an attractive approach for single-cell culture applications, due to its flexibility of adjustable cell culture spaces for different culture strategies, without decreasing isolation efficiency.

Here, we present a protocol for isolating and culturing single cells with a microfluidic platform, which utilizes a new microwell design concept to allow for high-efficiency single cell isolation and long-term clonal culture.

ومنصة ميكروفلويديك لعزل وحيدة الخلية عالية الإنتاجية والثقافة

Studying the heterogeneity of single cells is crucial for many biological questions, but is technically difficult. Thus, there is a need for a simple, yet ...

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.

A مجموعة القطيفة Femtoliter تخليق البروتين موازية على نطاق واسع من جزيئات الحمض النووي واحد

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

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.

متعدد خطوة الطباعة التصويرية متغير الطول لمحكم بصمام متعدد الأجهزة ميكروفلويديك

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

A Paired Bead and Magnet Array for Molding Microwells with Variable Concave Geometries

A spheroid culture is a useful tool for understanding cellular behavior in that it provides an in vivo-like three-dimensional environment. Various spheroid production methods such as non-adhesive surfaces, spinner flasks, hanging drops, and microwells have been used in studies of cell-to-cell interaction, immune-activation, drug screening, stem cell differentiation, and organoid generation. Among these methods, microwells with a three-dimensional concave geometry have gained the attention of scientists and engineers, given their advantages of uniform-sized spheroid generation and the ease with which the responses of individual spheroids can be monitored. Even though cost-effective methods such as the use of flexible membranes and ice lithography have been proposed, these techniques incur serious drawbacks such as difficulty in controlling the pattern sizes, achievement of high aspect ratios, and production of larger areas of microwells. To overcome these problems, we propose a robust method for fabricating concave microwells without the need for complex high-cost facilities. This method utilizes a 30 x 30 through-hole array, several hundred micrometer-order steel beads, and magnetic force to fabricate 900 microwells in a 3 cm x 3 cm polydimethylsiloxane (PDMS) substrate. To demonstrate the applicability of our method to cell biological applications, we cultured adipose stem cells for 3 days and successfully produced spheroids using our microwell platform. In addition, we performed a magnetostatic simulation to investigate the mechanism, whereby magnetic force was used to trap the steel beads in the through-holes. We believe that the proposed microwell fabrication method could be applied to many spheroid-based cellular studies such as drug screening, tissue regeneration, stem cell differentiation, and cancer metastasis.

This manuscript introduces a robust method of fabricating concave microwells without the need for complex high-cost facilities. Using magnetic force, steel beads, and a through-hole array, several hundred microwells were formed in a 3 cm x 3 cm polydimethylsiloxane (PDMS) substrate.