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. Author manuscript; available in PMC: 2018 Jan 1.
Published in final edited form as: Methods Mol Biol. 2017;1486:437–467. doi: 10.1007/978-1-4939-6421-5_17

Figure 6.

Figure 6

Lipid bilayer blocks nonspecific interactions while allowing kinetochores to be specifically anchored to the coverslip. (A) Kinetochores nonspecifically adsorbed on a clean glass surface at high density. Kinetochore components Mtw1 and Nuf2 that are fluorescently labeled are shown in red and green false colors, respectively. Image color channel registration is not corrected, so that the co-localized particles appear as red-green doublets. (B) The addition of 0.5 mg/ml κ-casein does not block nonspecific interaction to a workable level for single-molecule experiments. (C) A POPC lipid bilayer in coordination with 0.1 mg/ml κ-casein effectively blocks nonspecific adsorption of kinetochores onto the surface. (D) After specific linkers were provided to the same surface in (C), kinetochores were able to anchor specifically via their His-tags. Scale bar, 5 μm.