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Dual color localization microscopy of cellular nanostructures.

Dual color localization microscopy of cellular nanostructures.

Authors :
Gunkel M
Erdel F
Rippe K
Lemmer P
Kaufmann R
Hörmann C
Amberger R
Cremer C
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.

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