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Bőcskei-Antal Barnabás

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Bőcskei-Antal Barnabás
Super-resolution investigation of liposomal nanosystems

Aug 29 - kedd

15:30 – 17:00

I. Poszterszekció

P06

Super-resolution investigation of liposomal nanosystems

Barnabás Bőcskei-Antal, Ádám Orosz, György Török, Balázs Kiss

Semmelweis University, Budapest, Department of Biophysics and Radiation Biology

Dynamic light scattering is a method used for a long time in the study of nano-sized lipid vesicles (~100 nm in diameter). Up until now, no imaging methodology has been used that directly reveals the inner structure of vesicles in this size range. However, the use of a method that makes the structure of nanovesicles visible under physiological conditions would have significant advantages in the planning and development of therapeutic use. Several research teams have already imaged liposomes using confocal microscopy, but this microscopic method does not have a sufficiently high resolution to enable a clear separation of the membrane and the internal fluid space. Our aim was to develop a methodology that the structure of liposomal systems can be visualized in a high-resolution, robust and reproducible manner.

For our work, small unilamellar vesicles produced from DPPC molecules were labeled with fluorescent (Alexa Fluor 594) lipopolysaccharide molecules, and this system was examined by dynamic light scattering and STED microscopy.

The diameter of the liposomes extruded through the 100 nm pore opening was found to be almost the same both during the light scattering measurement and in the confocal reconstruction images (~100‑110 nm). After finding the right DPPC/LPS ratio and finding the optimal STED settings, we managed to record images with a good signal-to-noise ratio. Through the deconvolution processing of the STED images, the internal aqueous phase of the liposomes can also be successfully explored.

Our research group previously dealt with photoreactive damage of the membranes. To test our methodology, we also followed this process using our developed STED approach.

DPPC – Dipalmitoylphosphatidylcholine
LPS – Lipopolysaccharide
STED – Stimulated Emission Depletion