Work

Measuring the Size of Biological Nanostructures with Spatially Modulated Illumination Microscopy

Year: 2004

Type: article

Abstract: Spatially modulated illumination fluorescence microscopy can in theory measure the sizes of objects with a diameter ranging between 10 and 200 nm and has allowed accurate size measurement of subresolution fluorescent beads ( approximately 40-100 nm). Biological structures in this size range have so far been measured by electron microscopy. Here, we have labeled sites containing the active, hyperphosphorylated form of RNA polymerase II in the nucleus of HeLa cells by using the antibody H5. The spatially modulated illumination-microscope was compared with confocal laser scanning and electron microscopes and found to be suitable for measuring the size of cellular nanostructures in a biological setting. The hyperphosphorylated form of polymerase II was found in structures with a diameter of approximately 70 nm, well below the 200-nm resolution limit of standard fluorescence microscopes.

Source: Molecular Biology of the Cell

DOI: 10.1091/mbc.e04-01-0045

Authors: Sonya Martin, Antonio Virgilio Failla, Udo Spöri, Christoph Cremer, Ana Pombo

Institutions: Hammersmith Hospital, Imperial College London, Heidelberg University

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: European Molecular Biology Organization, Wellcome Trust, Deutsche Forschungsgemeinschaft, Medical Research Council

OpenAlex ID: W2162001304

FWCI: 4.598

Cites: 26

Cited by: 55

Related to: 10