Single-molecule localisation microscopy: accounting for chance co-localisation between foci in bacterial cells.

ABSTRACT: Using single-molecule fluorescence microscopes, individual biomolecules can be observed within live bacterial cells. Using differently coloured probes, physical associations between two different molecular species can be assessed through co-localisation measurements. However, bacterial cells are finite and small (~ 1 μm) relative to the resolution limit of optical microscopes (~ 0.25 μm). Furthermore, the images produced by optical microscopes are typically two-dimensional projections of three-dimensional objects. These limitations mean that a certain proportion of object pairs (molecules) will inevitably be assigned as being co-localised, even when they are distant at molecular distance scales (nm). What is this proportion? Here, we attack this problem, theoretically and computationally,

SUBMITTER: Aberg C 

PROVIDER: S-EPMC8448688 | biostudies-literature | 2021 Oct

REPOSITORIES: biostudies-literature