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Dual color localization microscopy of cellular nanostructures.
Dual color localization microscopy of cellular nanostructures.
- Source :
-
Biotechnology journal [Biotechnol J] 2009 Jun; Vol. 4 (6), pp. 927-38. - Publication Year :
- 2009
-
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.
- Subjects :
- Adenosine Triphosphatases genetics
Adenosine Triphosphatases metabolism
Cell Line, Tumor
Chromatin metabolism
Chromatin Assembly and Disassembly physiology
Chromosomal Proteins, Non-Histone genetics
Chromosomal Proteins, Non-Histone metabolism
Computer Simulation
Green Fluorescent Proteins genetics
Green Fluorescent Proteins metabolism
Histones genetics
Histones metabolism
Humans
Luminescent Proteins genetics
Luminescent Proteins metabolism
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins metabolism
Software
Red Fluorescent Protein
Cells ultrastructure
Chromatin ultrastructure
Image Processing, Computer-Assisted methods
Microscopy, Fluorescence methods
Subjects
Details
- Language :
- English
- ISSN :
- 1860-7314
- Volume :
- 4
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Biotechnology journal
- Publication Type :
- Academic Journal
- Accession number :
- 19548231
- Full Text :
- https://doi.org/10.1002/biot.200900005