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BioNanoScience

, Volume 8, Issue 1, pp 207–217 | Cite as

Easy and Fast Preparation of Large and Giant Vesicles from Highly Confined Thin Lipid Films Deposited at the Air–Water Interface

  • Valter Bavastrello
  • Adriano Caliari
  • Isabella Pesce
  • Luis A. Bagatolli
  • Martin M. Hanczyc
Article

Abstract

Lipid vesicles are supramolecular structures of great interest for industrial and research applications. They can be used simply to compartmentalize solutions and pack active molecules in femtoliter-scale volumes or as highly sophisticated drug delivery vehicles and dynamic cell-size bioreactors. For these reasons, many methods for the production of vesicles have been developed, and some of them present several drawbacks, such as long working times and the requirement of specific equipment to perform the technique. In this work, we present a method to produce vesicles from highly confined lipid films at the air–water interface. The procedure involves two simple steps: the formation of the thin lipid film at the air–water interface and then brief sonication (10 s). These films are obtained by depositing different aliquots of lipid organic solutions at the air–liquid interface of round-bottom Eppendorfs tubes. The morphology of the highly confined lipid thin films was studied by optical microscopy noting the formation of non-uniform depositions at the air–liquid interface, with the presence of thicker portions close to the container sidewall. Post-sonication, the presence of vesicles composed of 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC) or 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC) was confirmed using the complementary techniques of fluorescence microscopy and flow cytometry. The size distribution investigations carried out by flow cytometry revealed the optimal concentrations to favor the formation of giant vesicles (GVs). Furthermore, we investigated aqueous phase encapsulation by adding calcein or green fluorescent protein (GFP) to the aqueous phase then characterized by fluorescence microscopy and flow cytometry. We demonstrate a fast and easy method for producing vesicles including GVs on demand.

Keywords

Giant vesicles Langmuir films Phospholipids Encapsulation Protocol Flow cytometry Microscopy 

Notes

Acknowledgements

We would like to thank Prof Tetsuya Yomo for informative discussions. LAB wants to thank Program Prometeo, SENESCYT, Ecuador, for support.

Funding Information

V.B and M.M.H. were financially supported in part by the European Commission FP7 Future and Emerging Technologies Proactive (EVOBLISS 611640).

Supplementary material

12668_2017_464_MOESM1_ESM.doc (11.4 mb)
ESM 1 (DOC 11711 kb)

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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Valter Bavastrello
    • 1
  • Adriano Caliari
    • 1
  • Isabella Pesce
    • 1
  • Luis A. Bagatolli
    • 2
  • Martin M. Hanczyc
    • 1
    • 3
  1. 1.Centre for Integrative Biology (CIBIO)University of TrentoPovoItaly
  2. 2.Memphys-Center for Biomembrane Physics/Yachay EP and Yachay Tech UniversityUrcuquiEcuador
  3. 3.Chemical and Biological EngineeringUniversity of New MexicoAlbuquerqueUSA

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