Quantification of labelled target molecules via super-resolution dSTORM localization microscopy

Dániel Varga, Hajnalka Majoros, Zsuzsanna Ujfaludi, Tibor Pankotai, Miklós Erdélyi

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

The advent of super-resolution microscopy techniques has begun a revolution in the field of optical microscopy. Localization methods have become one of the most dynamically developed areas in this field. The one order of magnitude improvement in the lateral resolution has revealed a lot of new details mainly in the field of biology, which was hidden with conventional microscopes by the diffraction. However, the visualization of these measurements often go hand in hand with information loss and the quantification of such images become limited. Here we present a quantitative evaluation method which uses the raw localization coordinates and the associated precision for quantitative analysis. With our cluster analysis-based method we were able to determine the different properties of the selected clusters, such as their area (or in case of 3D images the volume), spatial distribution and the number of labelled target molecules in them, which is not follows directly from the reconstructed image due the stochastic nature of blinking and the often unknown labelling stoichiometry. The great advantage of our method is that we can gain the latter property from the sample directly. We also applied our method to investigate the repair mechanism of DNA double-strand breaks (DSBs).

Original languageEnglish
Title of host publicationSingle Molecule Spectroscopy and Superresolution Imaging XIII
EditorsIngo Gregor, Felix Koberling, Rainer Erdmann
PublisherSPIE
ISBN (Electronic)9781510632554
DOIs
Publication statusPublished - Jan 1 2020
EventSingle Molecule Spectroscopy and Superresolution Imaging XIII 2020 - San Francisco, United States
Duration: Feb 1 2020Feb 2 2020

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume11246
ISSN (Print)1605-7422

Conference

ConferenceSingle Molecule Spectroscopy and Superresolution Imaging XIII 2020
CountryUnited States
CitySan Francisco
Period2/1/202/2/20

Keywords

  • Chromatin organization
  • Cluster analysis
  • DNA DSB
  • Protein counting
  • Quantitative imaging
  • SMLM
  • Super-resolution microscopy
  • Trajectory fitting

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging

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  • Cite this

    Varga, D., Majoros, H., Ujfaludi, Z., Pankotai, T., & Erdélyi, M. (2020). Quantification of labelled target molecules via super-resolution dSTORM localization microscopy. In I. Gregor, F. Koberling, & R. Erdmann (Eds.), Single Molecule Spectroscopy and Superresolution Imaging XIII [1124612] (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 11246). SPIE. https://doi.org/10.1117/12.2545099