• Vesicle Prep Pro

    市场上第一款全自动制备无溶剂巨型单层囊泡(GUVs)的设备
  • Vesicle Prep Pro

    GUVs大小一致-适用与不同的应用

2018 - Method for immobilization of living and synthetic cells for high-resolution imaging and single-particle tracking

icon vpp   Vesicle Prep Pro publication in Scientific Reports (2018)

Authors:
Syga Ł., Spakman D., Punter C.M., Poolman B.

Journal:
Scientific Reports (2018) 13789


Abstract:

Super-resolution imaging and single-particle tracking require cells to be immobile as any movement reduces the resolution of the measurements. Here, we present a method based on APTES-glutaraldehyde coating of glass surfaces to immobilize cells without compromising their growth. Our method of immobilization is compatible with Saccharomyces cerevisiae, Escherichia coli, and synthetic cells (here, giant-unilamellar vesicles). The method introduces minimal background fluorescence and is suitable for imaging of single particles at high resolution. With S. cerevisiae we benchmarked the method against the commonly used concanavalin A approach. We show by total internal reflection fluorescence microscopy that modifying surfaces with ConA introduces artifacts close to the glass surface, which are not present when immobilizing with the APTES-glutaraldehyde method. We demonstrate validity of the method by measuring the diffusion of membrane proteins in yeast with single-particle tracking and of lipids in giant-unilamellar vesicles with fluorescence recovery after photobleaching. Importantly, the physical properties and shape of the fragile GUVs are not affected upon binding to APTES-glutaraldehyde coated glass. The APTES-glutaraldehyde is a generic method of immobilization that should work with any cell or synthetic system that has primary amines on the surface.


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