Dual color localization microscopy of cellular nanostructures.
Document Type
Article
Publication Date
2009
Keywords
Cell-Line-Tumor, Cells, Chromatin, Chromatin-Assembly-and-Disassembly, Chromosomal-Proteins-Non-Histone, Computer-Simulation, Green-Fluorescent-Proteins, Histones, Humans, Image-Processing-Computer-Assisted, Luminescent-Proteins, Microscopy-Fluorescence, Recombinant-Fusion-Proteins, Software
First Page
927
Last Page
938
JAX Location
see Reprint Collection (a pdf is available)
JAX Source
Biotechnol J 2009 Jun; 4(6):927-38.
Abstract
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.
Recommended Citation
Gunkel M,
Erdel F,
Rippe K,
Lemmer P,
Kaufmann R,
Hormann C,
Amberger R,
Cremer C.
Dual color localization microscopy of cellular nanostructures. Biotechnol J 2009 Jun; 4(6):927-38.