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Dual color localization microscopy of cellular nanostructures
Author(s) -
Gunkel Manuel,
Erdel Fabian,
Rippe Karsten,
Lemmer Paul,
Kaufmann Rainer,
Hörmann Christoph,
Amberger Roman,
Cremer Christoph
Publication year - 2009
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.200900005
Subject(s) - microscopy , super resolution microscopy , fluorescence microscope , fluorescence , biological system , resolution (logic) , nucleus , optical microscope , image resolution , microscope , biophysics , macromolecule , nanotechnology , optics , materials science , chemistry , physics , biology , computer science , scanning electron microscope , artificial intelligence , microbiology and biotechnology , biochemistry
The dual color localization microscopy (2CLM) presented here is based on the principles of spectral precision distance microscopy (SPDM) with conventional autofluorescent proteins under special physical conditions. This technique allows us to measure the spatial distribution of single fluorescently labeled molecules in entire cells with an effective optical resolution comparable to macromolecular dimensions. Here, we describe the application of the 2CLM approach to the simultaneous nanoimaging of cellular structures using two fluorochrome types distinguished by different fluorescence emission wavelengths. The capabilities of 2CLM for studying the spatial organization of the genome in the mammalian cell nucleus are demonstrated for the relative distributions of two chromosomal proteins labeled with autofluorescent GFP and mRFP1 domains. The 2CLM images revealed quantitative information on their spatial relationships down to length‐scales of 30 nm.