Nanostructure of specific chromatin regions and nuclear complexes.
Spatially modulated illumination (SMI) microscopy is a method of widefield fluorescence microscopy featuring interferometric illumination, which delivers structural information about nanoscale features in fluorescently labeled cells. Using this approach, structural changes in the context of gene activation and chromatin remodeling may be revealed. In this paper we present the application of SMI microscopy to size measurements of the 7q22 gene region, giving us a size estimate of 105+/-16 nm which corresponds to an average compaction ratio of 1:324. The results for the 7q22 domain are compared with the previously measured sizes of other fluorescently labeled gene regions, and to those obtained for transcription factories. The absence of a correlation between the measured and genomic sizes of the various gene regions indicate that a high variability in chromatin folding is present, with factors other than the sequence length contributing to the chromatin compaction. Measurements of the 7q22 region in different preparations and at different excitation wavelengths show a good agreement, thus demonstrating that the technique is robust when applied to biological samples.
Mathée
H
H
Applied Optics and Information Processing, Kirchhoff Institute für Physik, Universität Heidelberg, INF 227, 69120, Heidelberg, Germany.
Baddeley
D
D
Wotzlaw
C
C
Fandrey
J
J
Cremer
C
C
Birk
U
U
D002460:Cell Line; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000648:ultrastructure*; D002843:Chromatin / Q000648:ultrastructure*; D002875:Chromosomes / Q000235:genetics / Q000648:ultrastructure; D015925:Cryopreservation; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D008214:Lymphocytes / Q000648:ultrastructure; D014158:Transcription, Genetic
D002843:Chromatin