Revealing Spatial Molecular Heterogeneity of High-Density Biofunctionalized Surfaces Using DNA-PAINT.

ABSTRACT: The quantification and control of molecular densities and distributions on biofunctionalized surfaces are key for enabling reproducible functions in biosciences. Here, we describe an analysis methodology for quantifying the density and spatial distribution of high-density biofunctionalized surfaces, with densities in the order of 10²-10⁵ biomolecules per μm² area, in a short measurement time. The methodology is based on single-molecule DNA-PAINT imaging combined with simulation models that compensate for lifetime and spatial undersampling effects, resulting in three distinct molecule counting methods and a statistical test for spatial distribution. The analysis methodology is exemplified for a surface with ssDNA affinity binder molecules coupled to a PLL-

SUBMITTER: Tan WS 

PROVIDER: S-EPMC11533166 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature