A workflow for sizing oligomeric biomolecules based on cryo single molecule localization microscopy.

ABSTRACT: Single molecule localization microscopy (SMLM) has enormous potential for resolving subcellular structures below the diffraction limit of light microscopy: Localization precision in the low digit nanometer regime has been shown to be achievable. In order to record localization microscopy data, however, sample fixation is inevitable to prevent molecular motion during the rather long recording times of minutes up to hours. Eventually, it turns out that preservation of the sample's ultrastructure during fixation becomes the limiting factor. We propose here a workflow for data analysis, which is based on SMLM performed at cryogenic temperatures. Since molecular dipoles of the fluorophores are fixed at low temperatures, such an approach offers the possibility to use the orientation of the dipol

SUBMITTER: Schneider MC 

PROVIDER: S-EPMC7817001 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature