Membrane topography and the overestimation of protein clustering in single molecule localisation microscopy - identification and correction.

ABSTRACT: According to single-molecule localisation microscopy almost all plasma membrane proteins are clustered. We demonstrate that clusters can arise from variations in membrane topography where the local density of a randomly distributed membrane molecule to a degree matches the variations in the local amount of membrane. Further, we demonstrate that this false clustering can be differentiated from genuine clustering by using a membrane marker to report on local variations in the amount of membrane. In dual colour live cell single molecule localisation microscopy using the membrane probe DiI alongside either the transferrin receptor or the GPI-anchored protein CD59, we found that pair correlation analysis reported both proteins and DiI as being clustered, as did its derivative pair correlation-p

SUBMITTER: Adler J 

PROVIDER: S-EPMC11217499 | biostudies-literature | 2024 Jun

REPOSITORIES: biostudies-literature