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The potential of 3D‐FISH and super‐resolution structured illumination microscopy for studies of 3D nuclear architecture
Author(s) -
Markaki Yolanda,
Smeets Daniel,
Fiedler Susanne,
Schmid Volker J.,
Schermelleh Lothar,
Cremer Thomas,
Cremer Marion
Publication year - 2012
Publication title -
bioessays
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.175
H-Index - 184
eISSN - 1521-1878
pISSN - 0265-9247
DOI - 10.1002/bies.201100176
Subject(s) - chromatin , biology , fish <actinopterygii> , microscopy , fluorescence in situ hybridization , nuclear dna , dna , nuclear lamina , ultrastructure , evolutionary biology , biophysics , chromosome , anatomy , optics , gene , genetics , nuclear protein , physics , mitochondrial dna , transcription factor , fishery
Three‐dimensional structured illumination microscopy (3D‐SIM) has opened up new possibilities to study nuclear architecture at the ultrastructural level down to the ∼100 nm range. We present first results and assess the potential using 3D‐SIM in combination with 3D fluorescence in situ hybridization (3D‐FISH) for the topographical analysis of defined nuclear targets. Our study also deals with the concern that artifacts produced by FISH may counteract the gain in resolution. We address the topography of DAPI‐stained DNA in nuclei before and after 3D‐FISH, nuclear pores and the lamina, chromosome territories, chromatin domains, and individual gene loci. We also look at the replication patterns of chromocenters and the topographical relationship of Xist ‐RNA within the inactive X‐territory. These examples demonstrate that an appropriately adapted 3D‐FISH/3D‐SIM approach preserves key characteristics of the nuclear ultrastructure and that the gain in information obtained by 3D‐SIM yields new insights into the functional nuclear organization.