Name: De druppeloverdrachtsmethode gebruiken om op betrouwbare wijze gigantische unilamellaire blaasjes te bereiden (Video) | JoVE Journal
Uploaded: 2025-09-19T12:45:00.000+00:00
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Description: Leer de druppeloverdrachtsmethode om Giant Unilamellar Vesicles (GUV's) betrouwbaar te bereiden voor synthetische biologie en microrobotica. Dit protocol biedt hoge inkapselingsefficiëntie en is toegankelijk voor laboratoria met basisuitrusting.

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<li>Fujii, S., Matsuura, T., Sunami, T., Nishikawa, T., Kazuta, Y., Yomo, T. <a target="_blank" href="https://pubmed.ncbi.nlm.nih.gov/24901741/">Liposome display for in vitro selection and evolution of membrane proteins.</a> Nat Protoc. 9 (7), 1578-1591 (2014).</li>
<li>Matsushita-Ishiodori, Y., Hanczyc, M. M., Wang, A., Szostak, J. W., Yomo, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Using+imaging+flow+cytometry+to+quantify+and+optimize+giant+vesicle+production+by+water-in-oil+emulsion+transfer+methods%5BTitle%5D)+AND+%22Langmuir%22%5BJournal%5D)">Using imaging flow cytometry to quantify and optimize giant vesicle production by water-in-oil emulsion transfer methods.</a> Langmuir. 35 (6), 2375-2382 (2019).</li>
<li>Caliari, A., Hanczyc, M. M., Imai, M., Xu, J., Yomo, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Quantification+of+giant+unilamellar+vesicle+fusion+products+by+high-throughput+image+analysis%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">Quantification of giant unilamellar vesicle fusion products by high-throughput image analysis.</a> Int J Mol Sci. 24 (9), 8241(2023).</li>
<li>van Buren, L., Koenderink, G. H., Martinez-Torres, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((DisGUVery%3A+a+versatile+open-source+software+for+high-throughput+image+analysis+of+giant+unilamellar+vesicles%5BTitle%5D)+AND+%22ACS+Synth+Biol%22%5BJournal%5D)">DisGUVery: a versatile open-source software for high-throughput image analysis of giant unilamellar vesicles.</a> ACS Synth Biol. 12 (1), 120-135 (2023).</li>
<li>Pazzi, J., Subramaniam, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Nanoscale+curvature+promotes+high+yield+spontaneous+formation+of+cell-mimetic+giant+vesicles+on+nanocellulose+paper%5BTitle%5D)+AND+%22ACS+Appl+Mater+Interfaces%22%5BJournal%5D)">Nanoscale curvature promotes high yield spontaneous formation of cell-mimetic giant vesicles on nanocellulose paper.</a> ACS Appl Mater Interfaces. 12 (50), 56549-56561 (2020).</li>
<li>Ekosso, C., Liu, H., Glagovich, A., Nguyen, D., Maurer, S., Schrier, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Accelerating+the+discovery+of+abiotic+vesicles+with+AI-guided+automated+experimentation%5BTitle%5D)+AND+%22Langmuir%22%5BJournal%5D)">Accelerating the discovery of abiotic vesicles with AI-guided automated experimentation.</a> Langmuir. 41 (1), 858-867 (2025).</li>
<li>Lee, I. -H., Passaro, S., Ozturk, S., Ureña, J., Wang, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Intelligent+fluorescence+image+analysis+of+giant+unilamellar+vesicles+using+convolutional+neural+network%5BTitle%5D)+AND+%22BMC+Bioinformatics%22%5BJournal%5D)">Intelligent fluorescence image analysis of giant unilamellar vesicles using convolutional neural network.</a> BMC Bioinformatics. 23 (1), 48(2022).</li>
</ol>

<table><tbody><tr><td>0.22 &micro;m mixed cellulose esters membrane&nbsp; syringe filters</td><td>Merck</td><td>SLGSVR255F</td><td></td></tr><tr><td>1.5 mL tubes</td><td>Bioclass</td><td>06.0279.00</td><td></td></tr><tr><td>100 &mu;L glass syringe</td><td>Merck</td><td>20737</td><td></td></tr><tr><td>150 mL beaker</td><td>Avantor&nbsp;</td><td>213-1123</td><td>At least 3</td></tr><tr><td>20 mL dark glass vials ND24</td><td>Avantor&nbsp;</td><td>548-9001A</td><td></td></tr><tr><td>2 mL tubes</td><td>Bioclass</td><td>07.3534.99</td><td></td></tr><tr><td>50 mL cylinder</td><td>Avantor&nbsp;</td><td>612-3843</td><td></td></tr><tr><td>500 &mu;L glass syringe</td><td>Hamilton</td><td>549-1166</td><td></td></tr><tr><td>50 mL Falcon tube</td><td>Merck</td><td>CLS352070</td><td></td></tr><tr><td>50 mL Syringe&nbsp;</td><td>Fisher Scientific</td><td>8SS50L1</td><td></td></tr><tr><td>5 mL tubes</td><td>Greiner bio-one</td><td>622201</td><td></td></tr><tr><td>600 mL beaker</td><td>Avantor&nbsp;</td><td>213-1126</td><td></td></tr><tr><td>B&uuml;chner flask</td><td>Merck</td><td>Z740682</td><td></td></tr><tr><td>Cork rings for round bottom flasks</td><td>Avantor&nbsp;</td><td>217-1000</td><td></td></tr><tr><td>Crystallizer</td><td>Dutscher</td><td>080474A</td><td></td></tr><tr><td>D-(+)-Glucose</td><td>Sigma-Aldrich</td><td>G8270</td><td></td></tr><tr><td>D-(+)-Sucrose</td><td>Sigma-Aldrich</td><td>S9378</td><td></td></tr><tr><td>Digital thermo-hygrometer</td><td>Bohlender</td><td>V1863-07</td><td></td></tr><tr><td>DOPC</td><td>Avanti Research</td><td>850375C</td><td></td></tr><tr><td>Drill</td><td>Dremel</td><td>4000</td><td></td></tr><tr><td>Erioglaucine disodium salt (Brilliant Blue FCF)</td><td>Sigma-Aldrich</td><td>861146</td><td></td></tr><tr><td>G18 syringe needle</td><td>Terumo</td><td>AN1838R1</td><td></td></tr><tr><td>G21 syringe needle</td><td>Terumo</td><td>AN2138R1</td><td></td></tr><tr><td>Glove chamber</td><td>Bel-Art</td><td>F50030-0000</td><td>If available a glove box can be used but in laboratories with low humidity levels it may not be necessary</td></tr><tr><td>Hot plate magnetic stirrer</td><td>LLG Labware</td><td>6.263 448</td><td></td></tr><tr><td>Light mineral oil</td><td>Sigma-Aldrich</td><td>330779</td><td></td></tr><tr><td>New coccine</td><td>Sigma-Aldrich</td><td>199737</td><td></td></tr><tr><td>Nitrile Rubber O-Ring O-Ring, 8mm Bore, 12mm Outer Diameter</td><td>RS</td><td>196-4863</td><td></td></tr><tr><td>Open screw caps with septum ND24</td><td>Avantor&nbsp;</td><td>548-0031A</td><td></td></tr><tr><td>P1000 micropipette</td><td>Fisher Scientific</td><td>12346132</td><td></td></tr><tr><td>P10000 micropipette</td><td>Avantor&nbsp;</td><td>89079-978</td><td></td></tr><tr><td>P200 micropipette</td><td>Fisher Scientific</td><td>12326132</td><td></td></tr><tr><td>Paper towels</td><td>Avantor&nbsp;</td><td>KIMB6657</td><td></td></tr><tr><td>Parafilm</td><td>Avantor&nbsp;</td><td>291-1214</td><td></td></tr><tr><td>Pipette tips (1000)</td><td>Eppendorf</td><td>0030000919</td><td></td></tr><tr><td>Pipette tips (10000)</td><td>Eppendorf</td><td>0030000765</td><td></td></tr><tr><td>Pipette tips (200)</td><td>Eppendorf</td><td>0030000870</td><td></td></tr><tr><td>Polystyrene particles, 1.1 &micro;m</td><td>Sigma-Aldrich</td><td>LB11-1ML</td><td></td></tr><tr><td>Rubber cap</td><td>Bioclass S.r.l.</td><td>12.2300.05</td><td></td></tr><tr><td>Rubber conical gasket</td><td>Avantor&nbsp;</td><td>217-0211</td><td></td></tr><tr><td>Screw caps&nbsp; ND24</td><td>Avantor&nbsp;</td><td>548-0161A</td><td></td></tr><tr><td>Silicone oil AR20</td><td>Sigma-Aldrich</td><td>10836</td><td></td></tr><tr><td>Silicone pipe (internal diameter 1.6 mm external diameter 3 mm)</td><td>RS</td><td>667-8441</td><td></td></tr><tr><td>Silicone pipe (internal diameter 6 mm external diameter 12 mm)</td><td>RS</td><td>273-2541</td><td>At least 6 m</td></tr><tr><td>Three-ways junction</td><td>RS</td><td>795-405</td><td></td></tr><tr><td>Two-ways junction</td><td>RS</td><td>419-7287</td><td></td></tr><tr><td>Two-ways valve</td><td>Claber</td><td>91280</td><td></td></tr><tr><td>Ultrapure water</td><td>Thermo Fisher Scientific&nbsp;</td><td>22934</td><td>When available, this product can be replaced with water purified through Milli-Q systems</td></tr><tr><td>Universal Scissors</td><td>Bochem</td><td>4154</td><td></td></tr></tbody></table>

de druppeloverdrachtsmethode gebruiken om op betrouwbare wijze gigantische unilamellaire blaasjes te bereiden

Using the Droplet Transfer Method to Reliably Prepare Giant Unilamellar Vesicles

De druppeloverdrachtsmethode gebruiken om op betrouwbare wijze gigantische unilamellaire blaasjes te bereiden

Over the past two decades, the droplet transfer method, also known as inverted emulsion, double emulsion, phase transfer, or emulsion transfer, has proven ...

using-droplet-transfer-method-to-reliably-prepare-giant-unilamellar

Research

JoVE Journal

Bioengineering

Bio-engineering

Dit protocol beschrijft een werkwijze voor het bereiden extrusie lipide vesicles van sub-micron afmetingen met een hoge mate van homogeniteit. Deze methode maakt gebruik van een druk-gecontroleerd systeem met gecontroleerde stikstofstroom tarieven voor liposoom voorbereiding. De lipide...

Constant Pressure-gecontroleerde Extrusie werkwijze voor de bereiding van Nano-sized lipide bolletjes

Ondersteunde lipide dubbellagen en natuurlijke membraan deeltjes zijn handig systemen die de eigenschappen van celmembranen kunnen benaderen en diverse analytische strategieën worden opgenomen. Hier laten we een werkwijze voor het bereiden microarrays samengesteld ondersteunde lipide-bilaag beklede SiO2 parels, fosfolipide vesicula of natuurlijke membraan deeltjes.

Vorming van biomembraan Microarrays met een zuigmond-gebaseerde Vergadering Methode

Microfluïdische jetting tegen een druppel-interface lipide bilaag een betrouwbare manier om vesicles te genereren controle over membraan asymmetrie, incorporatie van transmembraan eiwitten en inkapseling materiaal. Deze techniek kan worden toegepast op een verscheidenheid van biologische systemen waarbij gecompartimenteerd biomoleculen gewenst bestuderen.

Lipidendubbellaag Vesicle Generation Met behulp Microfluidic Jetting

Reusachtige blaasjes die micrometergrote componenten bevatten, zijn nuttige celmodellen. De centrifugatiemethode voor water-in-olie-emulsie is een eenvoudige, krachtige tool voor de bereiding van reusachtige blaasjes met ingekapselde materialen.

Bereiding van Giant blaasjes inkapselen Microbolletjes door centrifugeren van een water-in-olie emulsie

Presenteren we een techniek voor contactloze micromanipulation van blaasjes, met behulp van de gelokaliseerde calcium ion verlopen. Microinjection van een calcium Ionenoplossing, in de nabijheid van een gigantische lipide blaasje, is gebruikt om het remodelleren van het lipide membraan, wat resulteert in de productie van membraan buisvormige uitsteeksels.

Membraan remodelleren van gigantische blaasjes in reactie op gelokaliseerde Calcium Ion verlopen

We demonstreren eencellige cultuur van bacteriën in gigantische blaasjes (GVS). Gv's met bacteriële cellen werden bereid door de druppel overdrachtsmethode en werden geïmmobiliseerd op een ondersteund membraan op een glas substraat voor directe observatie van bacteriële groei. Deze aanpak kan ook worden aangepast aan andere cellen.

Bacteriële celkweek op het eencellige niveau in gigantische blaasjes

Het doel van het protocol is om de mechanische eigenschappen van de membraan van gigantische blaasjes betrouwbaar te meten met micro pipet aspiratie.

Het verkrijgen van gigantische Unilamellaire hybride blaasjes door elektro vorming en meting van hun mechanische eigenschappen door micropipet aspiratie

Dit artikel introduceert een eenvoudige methode voor snelle productie van gigantische unilamellaire blaasjes met ingekapselde cytoskeletale eiwitten. De methode blijkt nuttig te zijn voor bottom-up reconstitutie van cytoskeletstructuren in opsluiting en cytoskelet-membraaninteracties.

Snelle inkapseling van gereconstitueerd cytoskelet in gigantische unilamellaire blaasjes

Dit artikel presenteert een methode voor het bereiden van samengestelde Giant Unilamellaire blaasjes (cGUV's) met een blaasje-in-blaasjesstructuur, met aangepaste elektrische geleidbaarheid in binnenste, ringvormige en buitenste gebieden. Elektroformatie synthetiseert eenvoudige GUV's, die via osmotische shock worden omgezet in stomatocyten en cGUV's, wat een waardevol model biedt voor het bestuderen van de biofysica van cellen met kern.

Synthese van samengestelde reusachtige unilamellaire blaasjes: een biomimetisch model van nucleaatcellen

Er wordt een experimenteel protocol gepresenteerd om gramicidine A-gedoopeerde druppelinterface-dubbellagen samen te stellen, elektrisch te stimuleren en te analyseren. Relaties tussen lipide-eiwitstructuur en -functie worden gekwantificeerd door veranderingen in membraanoppervlakte, ionische flux en enkelkanaalgeleidbaarheid te meten, en deze responsen te relateren aan plasticiteitsachtige veranderingen in ionische geleiding in een membraanmodel geïnspireerd door elektrische synapsen.

Lipid-eiwitmembraan structuur-functie karakterisering met behulp van druppelinterface-dubbellagen

Leer de druppeloverdrachtsmethode om Giant Unilamellar Vesicles (GUV's) betrouwbaar te bereiden voor synthetische biologie en microrobotica. Dit protocol biedt hoge inkapselingsefficiëntie en is toegankelijk voor laboratoria met basisuitrusting.

De druppeloverdrachtsmethode gebruiken om op betrouwbare wijze gigantische unilamellaire blaasjes te bereiden

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