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Liposomes pp 319–331Cite as

Environmental Scanning Electron Microscope Imaging of Vesicle Systems

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 606))

Abstract

The structural characteristics of liposomes have been widely investigated and there is certainly a strong understanding of their morphological characteristics. Imaging of these systems, using techniques such as freeze-fracturing methods, transmission electron microscopy, and cryo-electron imaging, has allowed us to appreciate their bilayer structures and factors that influence this. However, there are a few methods that study these systems in their natural hydrated state; commonly, the liposomes are visualized after drying, staining and/or fixation of the vesicles. Environmental scanning electron microscopy (ESEM) offers the ability to image a liposome in its hydrated state without the need for prior sample preparation. We were the first to use ESEM to study the liposomes and niosomes, and have been able to dynamically follow the hydration of lipid films and changes in liposome suspensions as water condenses onto, or evaporates from, the sample in real-time. This provides an insight into the resistance of liposomes to coalescence during dehydration, thereby providing an alternative assay for liposome formulation and stability.

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References

  1. Segal AW, Willis EJ, Richmond JE, Slavin G, Black CD, Gregoriadis G (1974) Morpho­logicalobservations on the cellular and subcellular destination of intravenously administered liposomes. Br J Exp Pathol 55:320-7

    CAS  PubMed  Google Scholar 

  2. Sternberg B, Sorgi FL, Huang L (1994) New structures in complex formation between DNA and cationic liposomes visualized by freeze—fracture electron microscopy. FEBS Lett 356:361-6

    Article  CAS  PubMed  Google Scholar 

  3. Xu Y, Hui S-W, Frederik P, Szoka FC Jr (1999) Pysicochemical characterisation and purification of cationic lipoplexes. Biophysical J 77:341-53

    Article  CAS  Google Scholar 

  4. Perrie Y, Frederik PM, Gregoriadis G (2001) Liposome-mediated immunisation: the effect of vesicle composition. Vaccine 19:3301-10

    Article  CAS  PubMed  Google Scholar 

  5. Davidsen J, Rosenkrands I, Christensen D, Vangala A, Kirby D, Perrie Y, Agger EM, Andersen P (2005) Characterization of cationic liposomes based on dimethyldioctadecylammonium and synthetic cord factor from M. tuberculosis (trehalose 6, 6′-dibehenate) - a novel adjuvant inducing both strong CMI and antibody responses. Biochim Biophys Acta 1718(1-2):22-31

    Article  CAS  PubMed  Google Scholar 

  6. Arunothayanun P, Turton JA, Uchegbu IF, Florence AT (1999) Preparation and in vitro/in vivo evaluation of luteinizing hormone releasing hormone (LHRH)-loaded polyhedral and spherical/tubular niosomes. J Pharm Sci 88:34-8

    Article  CAS  PubMed  Google Scholar 

  7. Mohammed AR, Weston N, Coombes AGA, Fitzgerald M, Perrie Y (2004) Liposome formulation of poorly water soluble drugs: optimisation of drug loading and ESEM analysis of stability. Int J Pharm 285:23-34

    Article  CAS  PubMed  Google Scholar 

  8. Donald AM (1998) Taking SEMs into a new environment. Mater World: 399-401

    Google Scholar 

  9. Egelhaaf RM, Epand RF, Maekawa S (2003) The arrangement of cholesterol in membranes and binding of NAP-22. Chem Phys Lipids 122:33-9

    Article  Google Scholar 

  10. McKinlay KJ, Allison FJ, Scotchford CA, Grant DM, Oliver JM, King JR, Wood JV, Brown PD (2004) Comparison of environmental scanning electron microscopy with high vacuum scanning electron microscopy as applied to the assessment of cell morphology. J Biomed Mater Res 69A:359-66

    Article  CAS  Google Scholar 

  11. Muscariello L, Rosso F, Marino G, Giordano A, Barbarisi M, Cafiero G, Barbarisi A (2005) A critical overview of ESEM applications in the biological field. J Cell Physiol 205:328-34

    Article  CAS  PubMed  Google Scholar 

  12. Robinson VN (1975) A wet stage modification to a scanning electron microscope. J Microsc 103:71-7

    CAS  PubMed  Google Scholar 

  13. Moncrieff DA, Robinson VNE, Harris LB (1978) Charge neutralisation of insulating surfaces in the SEM by gas ionisation. J Phys D: Appl Phys 11:2315-25

    Article  CAS  Google Scholar 

  14. Moncrieff DA, Barker PR, Robinson VNE (1979) Electron scattering by gas in the scanning electron microscope. J Phys D: Appl Phys 12:481-8

    Article  CAS  Google Scholar 

  15. Danilatos GD (1993) Introduction to the ESEM instrument. Microsc Res Tech 25:354-61

    Article  CAS  PubMed  Google Scholar 

  16. Danilatos GD (1990) Theory of the gaseous detector device in the ESEM. Adv Elect Electron Phy 78:1-102

    Google Scholar 

  17. Danilatos GD (1988) Foundations of environmental scanning electron microscope. Adv Elect Electron Phy 71:109-250

    CAS  Google Scholar 

  18. Donald AM, He C, Royall CP, Sferrazza M, Stelmashenko NA, Thiel BL (2000) Applications of environmental scanning electron microscopy to colloidal aggregation and film formation. Colloid Surface Physicochem Eng Aspect 174:37-53

    Article  CAS  Google Scholar 

  19. Stokes DJ (2003) Recent advances in electron imaging, image interpretation and applications: environmental scanning electron microscopy. Philos Transact A Math Phys Eng Sci 361:2771-87

    Article  CAS  PubMed  Google Scholar 

  20. Perrie Y, Mohammed AR, Vangala A, McNeil SE (2007) Environmental scanning electron microscopy studies of liposomes and niosomes. J Liposome Res 17(1):27-37

    Article  CAS  PubMed  Google Scholar 

  21. Vangala AK, Kirby D, Rosenkrands I, Agger E-M, Andersen P, Perrie Y (2006) A comparative study of cationic liposomes and niosome-based adjuvant systems for protein subunit vaccines Characterisation, Environmental Scan­ning Electron Microscopy analysis and immunisation studies. J Pharm Pharmacol 58:787-99

    Article  CAS  PubMed  Google Scholar 

  22. Elvira C, Fanovich A, Fernandez M, Fraile J, San RJ, Domingo C (2004) Evaluation of drug delivery characteristics of microspheres of PMMA-PCL-cholesterol obtained by supercritical-CO2 impregnation and by dissolution-evaporation techniques. J Control Release 99:231-40

    Article  CAS  PubMed  Google Scholar 

  23. D’Emanuele A, Dinarvand R (1995) Preparation, characterisation, and drug release from thermoresponsive microspheres. Int J Pharm 118:237-42

    Article  Google Scholar 

  24. Cao Y, Li H (2002) Interfacial activity of a novel family of polymeric surfactants. Eur Polymer J 38:1457-63

    Article  CAS  Google Scholar 

  25. Bangham AD, Standish MM, Watkins JC (1965) Diffusion of univalent ions across the lamellae of swollen phospholipids. J Mol Biol 13:328-52

    Google Scholar 

  26. Kirby C, Gregoriadis G (1984) Dehydration-rehydration vesicles: a simple method for high yield drug entrapment in liposomes. Biotechnology 2:979-84

    Article  CAS  Google Scholar 

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Acknowledgments

The authors gratefully acknowledge the financial support rendered: during the time of this work, Afzal Mohammed’s research was supported by Pfizer Global Research and The School of Life and Health Sciences, Aston University; Anil Vangala was awarded an Aston Scholarship; Daniel Kirby was funded by the European Commission (contract no. LSHP-CT-2003-503367), Habib Ali and Sarah McNeil were both funded through EPSRC Case awards, with additional support from Lipoxen Technologies Ltd for Sarah McNeil.

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Authors and Affiliations

  1. School of Life and Health Sciences, Aston University, Birmingham, UK

    Yvonne Perrie, Habib Ali, Daniel J. Kirby, Afzal U. R. Mohammed & Sarah E. McNeil

  2. School of Pharmacy and Chemistry, Kingston University, London, UK

    Anil Vangala

Authors
  1. Yvonne Perrie
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  2. Habib Ali
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  3. Daniel J. Kirby
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  4. Afzal U. R. Mohammed
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  5. Sarah E. McNeil
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  6. Anil Vangala
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Corresponding author

Correspondence to Yvonne Perrie .

Editor information

Editors and Affiliations

  1. College of Pharmacy, Midwestern University, N. 59th Avenue 19555, Glendale, 85308, U.S.A.

    Volkmar Weissig

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© 2010 Humana Press, a part of Springer Science+Business Media, LLC

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Perrie, Y., Ali, H., Kirby, D.J., Mohammed, A.U.R., McNeil, S.E., Vangala, A. (2010). Environmental Scanning Electron Microscope Imaging of Vesicle Systems. In: Weissig, V. (eds) Liposomes. Methods in Molecular Biology™, vol 606. Humana Press. https://doi.org/10.1007/978-1-60761-447-0_21

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  • DOI: https://doi.org/10.1007/978-1-60761-447-0_21

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-446-3

  • Online ISBN: 978-1-60761-447-0

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