title: Dual color localization microscopy of cellular nanostructures.
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.
authors: Kirchhoff-Institute for Physics & BioQuant Center University of Heidelberg, Heidelberg, Germany.
authors: Manuel
authors: M
authors: Gunkel
authors: Fabian
authors: F
authors: Erdel
authors: Karsten
authors: K
authors: Rippe
authors: Paul
authors: P
authors: Lemmer
authors: Rainer
authors: R
authors: Kaufmann
authors: Christoph
authors: C
authors: Hörmann
authors: Roman
authors: R
authors: Amberger
authors: Christoph
authors: C
authors: Cremer
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002477:Cells / Q000648:ultrastructure*; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure*; D042002:Chromatin Assembly and Disassembly / Q000502:physiology; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D003198:Computer Simulation; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012984:Software; D000097573:Red Fluorescent Protein
chemical_list: D000251:Adenosine Triphosphatases; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D049452:Green Fluorescent Proteins; D006657:Histones; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C468783:H2AX protein, human; C409462:SMARCA5 protein, human