A novel and robust method for counting components within bio-molecular complexes using fluorescence microscopy and statistical modelling.

ABSTRACT: Cellular biology occurs through myriad interactions between diverse molecular components, many of which assemble in to specific complexes. Various techniques can provide a qualitative survey of which components are found in a given complex. However, quantitative analysis of the absolute number of molecules within a complex (known as stoichiometry) remains challenging. Here we provide a novel method that combines fluorescence microscopy and statistical modelling to derive accurate molecular counts. We have devised a system in which batches of a given biomolecule are differentially labelled with spectrally distinct fluorescent dyes (label A or B), and mixed such that B-labelled molecules are vastly outnumbered by those with label A. Complexes, containing this component, are then simply score

SUBMITTER: Mersmann SF 

PROVIDER: S-EPMC9568568 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature