Single-Molecule Microscopy in the Life Sciences

Markus Axmann, Josef Madl, Gerhard J. Schütz

Research output: Chapter in Book/Report/Conference proceedingsChapterpeer-review

Abstract

Single-molecule microscopy is currently revolutionizing the life sciences. It appears to perfectly meet the demands of living matter: by singling out the individual entities, complexity becomes accessible to quantitative approaches. This chapter gives an overview of state-of-the-art single-molecule microscopy. We start by describing the potential of single-molecule methods in general: to follow unsynchronized processes, analyze heterogeneous samples, unravel structure-function relations, image nanostructures, and detect extremely rare events. Next, we provide the newcomer with a detailed discussion on technological requirements. Finally, we ask the questions: which parameters can we measure and what can they tell us in a biological experiment? We discuss the determination of single-molecule positions in three dimensions, orientation, and color. These parameters can be used to obtain insights into the association state of a biomolecule, its conformational state, or its mobility.

Original languageEnglish
Title of host publicationFluorescence Microscopy
Subtitle of host publicationFrom Principles to Biological Applications
PublisherWiley-VCH Verlag
Pages293-343
Number of pages51
ISBN (Print)9783527329229
DOIs
Publication statusPublished - 4 Apr 2013
Externally publishedYes

Keywords

  • Diffraction
  • Diffusion
  • Förster resonance energy transfer
  • Localization precision
  • Mean-square displacement
  • Mobility
  • Numerical aperture
  • Resolution
  • Single molecule biophysics

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