Nanostructure Analysis Using Spatially Modulated Illumination Microscopy

  • For an improved understanding of cellular processes, it is highly desirable to develop light optical methods for the analysis of biological nanostructures and their dynamics in the interior of three-dimensionally (3D) conserved cells. Here, important structural parameters to be considered are the topology, i.e. the mutual positions and distances, as well as the sizes of the constituting subunits. This has become possible by the development of a novel method of far-field light fluorescence microscopy, spatially modulated illumination (SMI) microscopy. Using this approach, axial distances between fluorescence-labeled targets can be measured with an accuracy close to 1 nm; their sizes can be determined down to a few tens of nanometers. This approach can be extended to the determination of 3D positions and mutual 3D distances and sizes of any number of small objects/subunits that can be discriminated due to their spectral signatures. Consequently, the new approach allows an ‘in situ nanostructure elucidation, until now regarded to be beyond the possibilities of far-field light microscopy. Application examples discussed are: colocalization/nanosizing and topological analysis of large protein-protein complexes, of nucleic acid-protein complexes (such as transcription factories), or of the highly complex DNA-protein nanostructures of which active/ inactive gene regions in the eukaryotic cell nucleus are constituted.

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Author:Antonio Virgillo Failla, Benno Albrecht, U. Spöri, A. Schweitzer, A. Kroll, Georg HildenbrandORCiD, M. Bach, Christoph Cremer
DOI:https://doi.org/https://doi.org/10.1159/000070464
ISSN:1424-8492
Parent Title (English):Complexus
Publisher:S. Karger AG
Document Type:Article
Language:English
Year of Completion:2003
Release Date:2024/11/25
GND Keyword:Fluoreszenzmikroskopie
Volume:1
Issue:2
Page Number:12
First Page:77
Last Page:88
Institutes:Fakultät Ingenieurwissenschaften und Informatik / Studiengänge / Medical Engineering and Data Science (Bachelor)
Research Focuses / Research Horizons:Materials / Material Testing & Sensor Technology
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