Name: controlled synthesis and fluorescence tracking of highly uniform poly(n-isopropylacrylamide) microgels
Uploaded: 2016-09-08T14:00:00
Description: Watch this Scientific Journal Video about Controlled Synthesis and Fluorescence Tracking of Highly Uniform PolyN-isopropylacrylamide Microgels at JoVE.com

The Deutsche Forschungsgemeinschaft (DFG) is acknowledged for financial support within the Sonderforschungsbereich SFB 985 "Functional Microgels and Microgel Systems".

1. microgel संश्लेषण नोट: एन -isopropylacrylamide (NIPAM) एन-हेक्सेन से recrystallized किया गया था। अन्य अभिकर्मकों के रूप में प्राप्त किया जाता था। <ol><li> पाली (NIPAM) मैट्रिक्स Microgels की परम्परागत बैच संश्लेषण <ol><li> 245 मिलीलीटर फ़िल...

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कार्यात्मक comonomer की थोड़ी मात्रा के अलावा कण आकार और PNIPAM निकाली गई microgels की संरचना पर एक महत्वपूर्ण प्रभाव हो सकता है। एक साथ छोटे पैमाने पर टेस्ट ट्यूब polymerization इस तरह के बदलाव ?...

उत्तेजनाओं के प्रति संवेदनशील पाली (एन -isopropylacrylamide) (PNIPAM) microgels 1,2 विभिन्न स्मार्ट अनुप्रयोगों में अपनी क्षमता के कारण पिछले दो दशकों में निरंतर ब्याज को आकर्षित किया है। प्रदर्शन उपयोग के मामलों switchable पायस स्टेबलाइजर्स 3-8, microlenses 9, आसान सेल कटाई 10,11 के लिए सेल संस्कृति substrates, और कम आणविक भार यौगिकों के लिए स्मार्ट वाहक और अन्य जैव चिकित्सा 12 उपयोगों में शामिल हैं। देखने का एक मौलिक अनुसंधान की दृष्टि से इन कणों ऐसे कोलाइडयन बातचीत 13-15 और बहुलक विलायक बातचीत 16-18 के रूप में विषयों की जांच के लिए उपयोगी साबित हो गया है। PNIPAM microgels और किसी भी दिए गए आवेदन में उनके डेरिवेटिव के सफल प्रयोग आम तौर पर मतलब कण आकार और कण आकार के वितरण की चौड़ाई पर ज्ञान की आवश्यकता है। PNIPAM सूक्ष्म शामिल प्रयोगात्मक परिणामों की सही व्याख्या के लिएजैल, कण संरचना है, जो कार्यात्मक comonomers से प्रभावित किया जा सकता है, के नाम से जाना पड़ता है। गतिशील और स्थिर प्रकाश बिखरने (DLS और एसएलएस, क्रमशः) विशिष्ट रूप से इस जानकारी को प्राप्त करने क्योंकि इन तरीकों तेजी से और प्रयोग करने के लिए अपेक्षाकृत आसान कर रहे हैं के लिए अनुकूल हैं; और वे अपने मूल पर्यावरण (फैलाव) में कण गुण जांच गैर invasively। डीएलएस और एसएलएस भी पूर्वाग्रह छोटा सा नमूना आकार, माइक्रोस्कोपी तरीकों के लिए ठेठ से उत्पन्न होने से बचने के कणों की विशाल संख्या से डेटा इकट्ठा। इसलिए, इस काम का पहला उद्देश्य कोलाइडयन लक्षण वर्णन करने के लिए नए चिकित्सकों के लिए प्रकाश बिखरने के बारे में अच्छा अभ्यास शुरू करने की है। आमतौर पर, वर्षा polymerization प्रयोगशाला पैमाने में किया जाता है और सही प्रतिक्रिया की स्थिति पाने के विशिष्ट गुण कण श्रमसाध्य हो सकता है और संश्लेषण के कई repetitions की आवश्यकता होती है सकते हैं के लिए किया जाता है। बड़ी खेप संश्लेषण के विपरीत, गैर हड़कंप मच वर्षा polymerization 19,20 एआर हैapid प्रक्रिया है जिसमें अलग-अलग अभिकारक रचना के बैच संकीर्ण आकार के वितरण के लिए एक साथ उपज कणों polymerized जा सकता है। एक साथ polymerization प्रयोगात्मक भिन्नता को कम करता है और बड़े उत्पादन का मतलब है सही प्रतिक्रिया की स्थिति प्रतिक्रिया बढ़ाने के लिए तेजी से पाया जा सकता है। इसलिए, हमारी दूसरी उद्देश्य प्रोटोटाइप में और अनुप्रयोगों है कि उत्पाद की एक बड़ी राशि की आवश्यकता नहीं है में गैर हड़कंप मच वर्षा polymerization की उपयोगिता का प्रदर्शन है। संश्लेषण और लक्षण के विभिन्न पहलुओं कोलाइडयन बातचीत अनुसंधान में फ्लोरोसेंट लेबल PNIPAM microgels के आवेदन के उदाहरण में एक साथ आते हैं। यहाँ हम एक विस्तृत मैट्रिक्स एकाग्रता रेंज पर लेबल हटाया गया मैट्रिक्स microgels के फैलाव में लेबल दरियाफ्त microgels के प्रसार की जांच और केंद्रित कोलाइडयन फैलाव में पिंजरे प्रभाव को हल करने के लिए बेहद सटीक एकल कण ट्रैकिंग का उपयोग करें। वाइड क्षेत्र प्रतिदीप्ति माइक्रोस्कोपी अच्छी तरह के लिए उपयुक्त हैआर इसे के रूप में इस उद्देश्य के संभावित विभिन्न मैट्रिक्स प्रजातियों में से एक बड़ी संख्या के बीच में कुछ दरियाफ्त अणुओं के विशिष्ट व्यवहार चिह्नित कर सकते हैं। इस तरह के डीएलएस, एसएलएस और rheology के रूप में तकनीक है, जो सिस्टम का पहनावा औसत गुणों को मापने और इसलिए एक बड़े प्रणाली में जांच के कणों की छोटी संख्या के व्यवहार को हल नहीं कर सकते हैं के विपरीत है। इसके अलावा, इस विशिष्ट उदाहरण में पारंपरिक प्रकाश बिखरने तरीकों उच्च कण एकाग्रता, जो मजबूत एकाधिक बिखरने किसी भी मानक विश्लेषण बेमानी करने के लिए सुराग के कारण भी उपयोग नहीं किया जा सकता। स्वचालित डाटा प्रोसेसिंग और सांख्यिकीय तरीकों का उपयोग समग्र प्रणाली एक कण ट्रैकिंग के लिए भी व्यवहार, जब बड़े नमूना आकार के औसत का विश्लेषण कर सकें।

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controlled synthesis and fluorescence tracking of highly uniform poly(n-isopropylacrylamide) microgels

उत्तेजनाओं के प्रति संवेदनशील पाली (एन -isopropylacrylamide) (PNIPAM) microgels विभिन्न संभावित व्यावहारिक आवेदन किया है और मौलिक अनुसंधान में उपयोग करता है। इस काम में, हम एक तेजी से गैर हड़कंप मच वर्षा polymerization प्रक्रिया द्वारा ट्यूनिंग microgel आकार के लिए एक प्रदर्शन के रूप में की fluorescently लेबल PNIPAM microgels पर नज़र रखने के एक कण का उपयोग करें। यह दृष्टिकोण अच्छी तरह से नए प्रतिक्रिया रचनाओं और शर्तों प्रोटोटाइप के लिए या अनुप्रयोगों है कि उत्पाद की बड़ी मात्रा की आवश्यकता नहीं है के लिए अनुकूल है। Microgel संश्लेषण, कण आकार और गतिशील और स्थिर प्रकाश बिखरने से संरचना निर्धारण प्रोटोकॉल में विस्तृत रहे हैं। यह दिखाया गया है कार्यात्मक comonomers के अलावा कण nucleation और संरचना पर एक बड़ा प्रभाव हो सकता है। व्यापक क्षेत्र प्रतिदीप्ति माइक्रोस्कोपी द्वारा एक कण ट्रैकिंग गैर लेबल microgels का एक केंद्रित मैट्रिक्स में लेबल दरियाफ्त microgels के प्रसार की एक जांच के लिए अनुमति देता है, एक सिस्टम को आसानी से जांच नहीं कीइस तरह के गतिशील प्रकाश बिखरने के रूप में अन्य तरीकों।

बैच में PNIPAM microgel कणों की संख्या, और इस तरह अंतिम कण की मात्रा, nucleation चरण 20 जल विरोधी सह मोनोमर डाई methacryloxyethyl thiocarbamoyl rhodamine बी बैच में कण संख्या घनत्व को कम करने से nucleation प्रभावित क...

Watch this Scientific Journal Video about Controlled Synthesis and Fluorescence Tracking of Highly Uniform PolyN-isopropylacrylamide Microgels at JoVE.com

Controlled Synthesis and Fluorescence Tracking of Highly Uniform PolyN-isopropylacrylamide Microgels

गैर-हड़कंप मच वर्षा polymerization उत्तेजनाओं के प्रति संवेदनशील पाली (एन -isopropylacrylamide) संकीर्ण आकार के वितरण के microgels के संश्लेषण के लिए एक तेजी से, प्रतिलिपि प्रस्तुत करने योग्य प्रोटोटाइप दृष्टिकोण प्रदान करता है। इस प्रोटोकॉल के संश्लेषण में, प्रकाश बिखरने लक्षण वर्णन है और एक व्यापक क्षेत्र माइक्रोस्कोपी सेटअप में इन microgels का एक कण प्रतिदीप्ति ट्रैकिंग प्रदर्शन कर रहे हैं।

नियंत्रित संश्लेषण और अत्यधिक वर्दी पाली की प्रतिदीप्ति ट्रैकिंग (<em&gt; एन</em&gt; -isopropylacrylamide) Microgels

Stimuli-sensitive poly(N-isopropylacrylamide) (PNIPAM) microgels have various prospective practical applications and uses in fundamental research. In this ...

The overall goal of this work is to investigate the dynamic behavior and interaction of soft colloidal particles in dispersions of various particle densities in the vicinity of glass transition. Microgels are soft objects that can adapt their size, shape, and mobility according to their environments. In contrast to rigid colloidal particles, there are many open questions concerning the behavior of soft microgels.The main advantage of wide field fluorescence microscopy is the possibility to localize single microgels with high accuracy, directly follow their motion in C2, and to analyze the amounts of diffusion. To investigate the dynamics of single microgels in dense systems, only a small number of microgels must be fluorescently labeled. Non-stirred precipitation polymerization allows for parallel synthesis of microgel with variable reaction conditions to conveniently match the size of labeled and unlabeled microgels.Dynamic light-scattering provides the average hydrodynamic radii of the microgels within an accuracy of a few nanometers. Dissolve 1.8 grams of NIPAM and 24 milligrams of BIS in 245 milliliters of filtered double-distilled water in a 500 milliliter, three-neck round bottom flask equipped with reflux condenser, a stirrer, and a rubber septum. Insert a thermometer and a 120-millimeter needle for the nitrogen input through the septum.Heat the solution to 60 degrees Celsius while stirring. Then, deoxygenate the solution by purging with nitrogen for 40 minutes. Simultaneously prepare an initiator solution of 155 milligrams of potassium persulfate, or KPS, in five milliliters of filtered double-distilled water, and bubble the solution with nitrogen to remove oxygen.Next, wash a syringe with nitrogen, and transfer the complete five milliliter KPS solution to a nitrogen-washed syringe equipped with a 120-millimeter needle. Lift the nitrogen needle above the solution level in the three-neck flask, and add the KPS solution rapidly through the rubber septum into the reactor. Let the polymerization proceed for one hour under nitrogen flow and slow stirring at 60 degrees Celsius.Use a Buchner funnel and filter paper to filter the hot reaction solution, and discard any big aggregates. After letting the dispersion cool down, centrifuge and redisperse the dispersion three times for 40 minutes at 257, 000 times G.Finally, redisperse the sediment in a minimum viable amount of double-distilled water before lyophilizing the dispersion for storing. Weigh 257.7 milligrams of NIPAM, 3.5 milligrams of BIS, and 1.5 milligrams of methacryloxyethyl thiocarbamoyl rhodamine B dye in a glass vessel, and add 10 milliliters of filter double-distilled water.Prepare the same solution without the dye in a separate glass vessel. Ultrasonicate the dye solution for 15 minutes to dissolve the dye in water. Prepare various dilutions of the monomer solution with the dye to obtain a concentration series.Here, dye in the concentration range of 02 to 0.1 millimole per liter is used. Next, dissolve 8.4 milligrams of KPS in 10 milliliters of filtered double-distilled water to get the initiator solution. Transfer 0.5 milliliters of the concentration series and 05 milliliters of the KPS solution to test tubes with a 10 millimeter diameter to obtain the final reaction solutions.Seal the tubes with rubber septa. Preheat an oil bath in a double-walled glass vessel connected to a heating circulator to 63 degrees Celsius. Deoxygenize the reaction solutions by purging with nitrogen through 120-millimeter needles for 20 minutes.Then, insert the tubes into a floating platform, and immerse the platform in the pre-heated oil bath. Set the temperature to 60 degrees Celsius. For high-precision particle size tuning, the temperature control during the initial reaction has to be rigorous, typically plus or minus 0.1 degrees Celsius.Let the reaction proceed for an appropriate amount of time. Typically, one hour is enough. Following the reaction, transfer the reaction tubes rapidly to an oven at 60 degrees Celsius.Put one drop of the hot dispersion in 10 milliliters of filtered double-distilled water, pre-heated over the poly-NIPAM volume phase transition temperature in order to measure the hydrodynamic radii of the particles in the collapsed state. Let the rest of the dispersions cool down to room temperature, and transfer them into centrifuge tubes. After centrifuging the solution, dilute the microgels in two milliliters of filtered double-distilled water to use as tracer particles.For hydrodynamic radius determination in the collapsed state by dynamic light scattering, first wash the cuvettes and glassware with acetone vapor. Transfer approximately one milliliter of diluted particle dispersion to a measurement cuvette. Temper the dynamic light-scattering goniometer index match bath to 50 degrees Celsius, and transfer the sample to the instrument without letting it cool down.Insert the sample cuvette, and move the detector arm to the small scattering angle. After checking the beam profile and count range as described in the text protocol, move the goniometer arm to the highest scattering angle. Check that the count rate is still high enough for the measurement.If the intensity is too low, move the arm to a lower scattering angle. Then, check the beam visually through the toluene bath glass at the lowest scattering angle. If glow around the incident beam is observed, multiple scattering takes place.Acquire 20 correlograms between the minimum and maximum scattering angle with a minimum acquisition time of 60 seconds. Increase the acquisition time for weak intensity large-scattering angles if necessary. Use an appropriate objective lens of the desired magnification and aperture for excitation of the tracers and simultaneous fluorescence collection from the sample.In this work, a 100X, 1.3 numerical aperture oil-immersion objective lens is used. Place the moisture chamber onto an XYZ pi-SO table that fits into a microscope. To prevent the sample from drying, place a plasma-cleaned cover slip into the moisture chamber, and pipette 10 microliters of the desired concentration of poly-NIPAM dispersion.Depending on the excitation and emission spectra of the fluorescent dye, use a suitable laser for excitation, and adjust the laser power appropriately. A 561-nanometer diode-pump solid state laser is used here at a constant laser power of 16 milliwatts. To obtain homogeneous sample illumination, couple the laser into a multimode fiber, shake the fiber using a vortexer to temporarily average out laser speckles, and project the fiber end into the sample plane.Calibrate the Z-distance from the back reflection of the cover slip, and focus several micrometers into the sample by moving the objective slightly up, and fix the Z-position using a Z-compensator. This avoids any interface effects with the cover slip. Adjust the detector parameters, such as exposure time, to the strength of the fluorescent signal.In this case, an EMCCD camera is used with an exposure time of 0.1 seconds in electron-multiplying mode and a gain of 50. Acquire several movies with the appropriate number of frames to obtain adequate lag time to calculate the mean-squared displacement of the microgels in different regions of the sample. Eight labeled microgel batches were synthesized.For six batches, correlogram decay rates linear rise against the square of the scattering vector magnitude, which indicates narrow size distribution. A fit through the origin indicates only translational diffusion. For two batches, nonlinear behavior is observed.The deviation is caused by broad size distribution and a particle form factor minimum, which coincides with the scattering vector range. To obtain a better estimate for the mean diffusion coefficient of the particles, exclude these points from the fit. High signal to noise ratio enables tracking of fluorescent microgels.Their diffusion is restricted by the the unlabeled microgel matrix. Higher concentrations of unlabeled microgel matrix on the right leads to a pronounced cage effect, where the fluorescent microgels are trapped in an unlabeled matrix of microgels. At low concentrations of unlabeled mircogel matrix, the tracer microgels exhibit normal diffusion, which is inferred from the linear increase of mean-square displacement with lag time.However, at high concentrations close to the colloidal glass transition, they exhibit nonlinear diffusion, depicted by nonlinear temporal evolution of mean-square displacements. At higher concentration of unlabeled microgel matrix, the tracer microgels are trapped by unlabeled neighbors into transient cages, as illustrated by clustered tracks for 12 microgels. Parallel small-scale synthesis enables fast experimentation with different reaction parameters.This also minimizes any undesired experimental variation due to reaction temperature differences. In combination with the careful dynamic light-scattering characterization, the reaction conditions for the right microgel size can be found fast. In this video, we demonstrate how controlled microgel synthesis allows us to investigate the diffusion of single microgels in concentrated microgel solutions by wide field fluorescence microscopy.Following this procedure, it will be possible to understand how the structure of microgels controls their properties. This will open the door to designing new soft interactive matter.

Research

JoVE Journal

Chemistry

Controlled Synthesis and Fluorescence Tracking of Highly Uniform Poly(N-isopropylacrylamide) Microgels

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

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

पाली के संश्लेषण (<em&gt; एन</em&gt; -isopropylacrylamide) एनिस्ट्रोपिक थर्मो-जवाबदेही और organophilic / हाइड्रोफिलिक लोड हो रहा है क्षमता के लिए जानूस Microhydrogels

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

Highly Stereoselective Synthesis of 1,6-Ketoesters Mediated by Ionic Liquids: A Three-component Reaction Enabling Rapid Access to a New Class of Low Molecular Weight Gelators

ईओण का तरल पदार्थ द्वारा मध्यस्थता 1,6-Ketoesters की अत्यधिक stereoselective संश्लेषण: कम आणविक भार Gelators का एक नया वर्ग के लिए तेजी से पहुँच सक्रिय तीन घटक रिएक्शन

In organic chemistry ionic liquids (ILs) have emerged as safe and recyclable reaction solvents. In the presence of a base ILs can be deprotonated to form ...

A Simple Method for the Size Controlled Synthesis of Stable Oligomeric Clusters of Gold Nanoparticles under Ambient Conditions

परिवेश की स्थिति के तहत सोने के नैनोकणों स्थिर Oligomeric क्लस्टर का आकार नियंत्रित संश्लेषण के लिए एक सरल विधि

Reducing dilute aqueous HAuCl4 with sodium thiocyanate (NaSCN) under alkaline conditions produces 2 to 3 nm diameter nanoparticles. Stable grape-like ...

Synthesis and Characterization of Supramolecular Colloids

संश्लेषण और supramolecular Colloids की विशेषता

Control over colloidal assembly is of utmost importance for the development of functional colloidal materials with tailored structural and mechanical ...

Hydroquinone Based Synthesis of Gold Nanorods

सोना nanorods की Hydroquinone आधारित संश्लेषण

Gold nanorods are an important kind of nanoparticles characterized by peculiar plasmonic properties. Despite their widespread use in nanotechnology, the ...

Scale-up Chemical Synthesis of Thermally-activated Delayed Fluorescence Emitters Based on the Dibenzothiophene-S,S-Dioxide Core

Dibenzothiophene-s, s-डाइऑक्साइड कोर के आधार पर स्केल-अप थर्मल-सक्रिय देरी प्रतिदीप्ति उत्सर्जक के रासायनिक संश्लेषण

We report a procedure to linearly scale-up the synthesis of 2,8-bis(3,6-di-tert-butyl-9H-carbazol-9-yl)dibenzothiophene-S,S-dioxide (compound 4) and ...

Synthesis and Mass Spectrometry Analysis of Oligo-peptoids

Oligo-peptoids का संश्लेषण एवं मास स्पेक्ट्रोमेट्री विश्लेषण

Peptoids are sequence-controlled peptide-mimicking oligomers consisting of N-alkylated glycine units. Among many potential applications, peptoids have ...

Synthesis of Substrate-Bound Au Nanowires Via an Active Surface Growth Mechanism

एक सक्रिय सतह वृद्धि तंत्र के माध्यम से सब्सट्रेट-बाउंड Au Nanowires का संश्लेषण

Advancing synthetic capabilities is important for the development of nanoscience and nanotechnology. The synthesis of nanowires has always been a ...

Synthesis and Characterization of Amphiphilic Gold Nanoparticles

एम्फीफिलिक गोल्ड नैनोकणों का संश्लेषण और विशेषता

Gold nanoparticles covered with a mixture of 1-octanethiol (OT) and 11-mercapto-1-undecane sulfonic acid (MUS) have been extensively studied because of ...

Hierarchical and Programmable One-Pot Oligosaccharide Synthesis

पदानुक्रमिक और प्रोग्रामेबल वन-पोट ओलिगोसैकराइड संश्लेषण

This article presents a general experimental protocol for programmable one-pot oligosaccharide synthesis and demonstrates how to use Auto-CHO software for ...