Skip to main content
SpringerLink
Log in
Book cover

Advanced Protocols in Oxidative Stress III pp 111–121Cite as

Evaluation of Physical Integrity of Lipid Bilayer Under Oxidative Stress: Application of Fluorescence Microscopy and Digital Image Processing

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1208))

Abstract

Membrane damage as a result of oxidative stress is quantified using digital image heterogeneity analysis of single giant unilamellar vesicles (GUVs) composed of soy phosphatidylcholine (PC), which were found to undergo budding when containing chlorophyll a (Chla) as photosensitizer in the lipid bilayer. Based on digital image heterogeneity analysis, a dimensionless scalar parameter “entropy” for the budding process was found to change linearly during an initial budding stage. Photo-induced peroxidation of PC to form linoleoyl hydroperoxides, further leading to domains of higher polarities in GUVs, was suggested to initiate the budding process. The effect on budding process of GUVs was suggested for use in assays for evaluation of potential protectors of lipid bilayer integrity under oxidative stress, and “entropy” seemed to be a valid descriptor of such membranal integrity. The one-step procedure for quantification of prooxidative effects and antioxidative protection provided by drug candidates and potential food ingredients in membranes could be easily automated for direct measurement of oxidative and antioxidative effects on cellular integrity.

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Halliwell B (2007) Oxidative stress and cancer: have we moved forward? Biochem J 401:1–11

    Article  PubMed  CAS  Google Scholar 

  2. Singh N, Dhalla AK, Seneviratne C, Singal PK (1995) Oxidative stress and heart failure. Mol Cell Biochem 147:77–81

    Article  PubMed  CAS  Google Scholar 

  3. Amer J, Ghoti H, Rachmilewitz E, Koren A, Levin C, Fibach E (2006) Red blood cells, platelets, and polymorphonuclear neutrophils of patients with sickle cell disease exhibit oxidative stress that can be ameliorated by antioxidants. Br J Haematol 132:108–113

    Article  PubMed  CAS  Google Scholar 

  4. Bangham AD, Horne RW (1964) Negative staining of phospholipids and their structural modifications by surface-active agents as observed in the electron microscope. J Mol Biol 8:660–668

    Article  PubMed  CAS  Google Scholar 

  5. Bangham AD, Horne RW, Glauert AM, Dingle JT, Lucy JA (1962) Action of saponin on biological cell membranes. Nature 196:952–955

    Article  PubMed  CAS  Google Scholar 

  6. Horne RW, Bangham AD, Whittaker VP (1963) Negatively stained lipoprotein membranes. Nature 200:1340–1340

    Article  PubMed  CAS  Google Scholar 

  7. Reeves JP, Dowben MW (1969) Formation and properties of thin-walled phospholipid vesicles. J Cell Physiol 73:49–60

    Article  PubMed  CAS  Google Scholar 

  8. Angelova MI, Soléau S, Meléard P, Faucon JF, Bothorel P (1992) Preparation of giant vesicles by external AC fields. Kinetics and application. Progr Colloid Polymer Sci 89:127–131

    Article  CAS  Google Scholar 

  9. Angelova MI, Dimitrov DS (1986) Liposome electroformation. Faraday Discuss Chem Soc 81:303–311

    Article  CAS  Google Scholar 

  10. Torchilin VP (2006) Multifunctional nanocarriers. Adv Drug Deliv Rev 58:1532–1555

    Article  PubMed  CAS  Google Scholar 

  11. Dutta D, Pulsipher A, Luo W, Mak H, Yousaf MN (2011) Engineering cell surfaces via liposome fusion. Bioconjug Chem 22:2423–2433

    Article  PubMed  CAS  Google Scholar 

  12. Dutta D, Pulsipher A, Luo W, Yousaf MN (2011) Synthetic chemoselective rewiring of cell structures: generation of three-dimensional tissue structures. J Am Chem Soc 133:8704–8713

    Article  PubMed  CAS  Google Scholar 

  13. Hernandez VA, Karlsson G, Edwards K (2011) Intrinsic heterogeneity in liposome suspensions caused by the dynamic spontaneous formation of hydrophobic active site in lipid membranes. Langmuir 27:4873–4883

    Article  Google Scholar 

  14. Menger FM, Gabrielson KD (1995) Cytomimetic organic chemistry: early developments. Angew Chem Int Ed Engl 34:2091–2106

    Article  CAS  Google Scholar 

  15. Menger FM, Keiper JS (1998) Chemistry and physics of giant vesicles as biomembrane models. Curr Opin Chem Biol 2:726–732

    Article  PubMed  CAS  Google Scholar 

  16. Menger FM, Azov VA (2000) Cytomimetic modeling in which one phospholipid liposome chemically attacks another. J Am Chem Soc 122:6492–6493

    Article  CAS  Google Scholar 

  17. Bagatolli LA (2006) To see or not to see: lateral organization of biological membranes and fluorescence microscopy. Biochim Biophys Acta 1758:1541–1556

    Article  PubMed  CAS  Google Scholar 

  18. Stottrup BL, Nguyen AH, Tüzel E (2010) Taking another look with fluorescence microscopy: image processing techniques in Langmuir monolayers for the twenty-first century. Biochim Biophys Acta 1798:1289–1300

    Article  PubMed  CAS  Google Scholar 

  19. Morales-Penningston NF, Wu J, Farkas ER, Goh SL, Konyakhina TM, Zheng JY, Webb WW, Feigenson GW (2010) GUV preparation and imaging: minimizing artifacts. Biochim Biophys Acta 1798:1324–1332

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  20. Haverstick DM, Glaster M (1987) Visualization of Ca2+-induced phospholipid domains. Proc Natl Acad Sci U S A 84:4475–4479

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  21. Haverstick DM, Glaster M (1988) Visualization of domain formation in the inner and outer leaflets of a phospholipid bilayer. J Cell Biol 106:1885–1892

    Article  PubMed  CAS  Google Scholar 

  22. Zhao J, Wu J, Shao H, Kong F, Jain N, Hunt G, Feigenson G (2007) Phase studies of model biomembranes: macroscopic coexistence of Lα + Lβ, with light-induced coexistence of Lα + Lo phases. Biochim Biophys Acta 1768:2777–2786

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  23. Heuvingh J, Bonneau S (2009) Asymmetric oxidation of giant vesicles triggers curvature-associated shape transition and permeabilization. Biophys J 97:2904–2912

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  24. Ayuyan AG, Cohen FS (2006) Lipid peroxides promote large rifts: effects of excitation of probes in fluorescence microscopy and electrochemical reactions during vesicle formation. Biophys J 91:2172–2183

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  25. Ishii K, Hamada T, Hatakeyama M, Sugimoto R, Nagasaki T, Takagi M (2009) Reversible control of exo- and endo-budding transitions in a photosensitive lipid membrane. Chembiochem 10:251–256

    Article  PubMed  CAS  Google Scholar 

  26. Choe E, Min DB (2006) Chemistry and reactions of reactive oxygen species in foods. Crit Rev Food Sci Nutr 46:1–22

    Article  PubMed  CAS  Google Scholar 

  27. Farge E, Devaux PF (1992) Shape changes of giant liposomes induced by an asymmetric transmembrane distribution of phospholipids. Biophys J 61:347–357

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  28. Gonzalez RC, Woods RE, Eddins SL (2003) Digital image processing using MATLAB. Prentice Hall, New Jersey, Chapter 11, pp 426–483

    Google Scholar 

  29. Moscho A, Orwar O, Chiu DT, Modi BP, Zare RN (1996) Rapid preparation of giant unilamellar vesicles. Proc Natl Acad Sci U S A 93:11443–11447

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  30. Liang R, Liu Y, Fu LM, Ai XC, Zhang JP, Skibsted LH (2012) Antioxidants and physical integrity of lipid bilayers under oxidative stress. J Agric Food Chem 60:10331–10336

    Article  PubMed  CAS  Google Scholar 

  31. Klein RA (1970) The detection of oxidation in liposome preparations. Biochim Biophys Acta 210:486–489

    Article  PubMed  CAS  Google Scholar 

  32. Naguib YMA (1998) A fluorometric method for measurement of peroxyl radical scavenging activities of lipophilic antioxidants. Anal Biochem 265:290–298

    Article  PubMed  CAS  Google Scholar 

  33. Naguib YMA (2000) A fluorometric method for measurement of oxygen radical-scavenging activity of water-soluble antioxidants. Anal Biochem 284:93–98

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work has been supported by the Natural Science Foundation of China (20803091). L.H.S. is grateful for support from the Danish Research Council for Technology and Production as the grant 09-065906/FTP: Redox communication in the digestive tract.

Author information

Authors and Affiliations

  1. Department of Chemistry, Renmin University of China, Beijing, 100872, China

    Ran Liang & Jian-Ping Zhang

  2. Food Chemistry, Department of Food Science, University of Copenhagen, Rolighedsvej 30, 1958, Frederiksberg C, Denmark

    Leif H. Skibsted

Authors
  1. Ran Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jian-Ping Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Leif H. Skibsted
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Leif H. Skibsted .

Editor information

Editors and Affiliations

  1. University of Florida, Gainsville, Florida, USA

    Donald Armstrong

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer Science+Business Media New York

About this protocol

Cite this protocol

Liang, R., Zhang, JP., Skibsted, L.H. (2015). Evaluation of Physical Integrity of Lipid Bilayer Under Oxidative Stress: Application of Fluorescence Microscopy and Digital Image Processing. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress III. Methods in Molecular Biology, vol 1208. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1441-8_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-1441-8_9

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1440-1

  • Online ISBN: 978-1-4939-1441-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation