Dual color localization microscopy of cellular nanostructures

Manuel Gunkel (1) , Fabian Erdel (2) , Karsten Rippe (2) , Paul Lemmer (1) , Rainer Kaufmann (1) , Christoph Hörmann (1) , Roman Amberger (1) , Christoph Cremer (1)

1 Kirchhoff Institut für Physik
2 Research Group Genome Organization and Function

The Dual Color Localization Microscopy (2CLM) presented here is based on the principles of Spectral Precision Distance Microscopy (SPDM) with conventional fluorochromes 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.

Mots clés: Life Sciences

Manuel Gunkel, Fabian Erdel, Karsten Rippe, Paul Lemmer, Rainer Kaufmann, et al.. Dual color localization microscopy of cellular nanostructures. Biotechnology Journal, 2009, 4 (6), pp.927. ⟨10.1002/biot.200900005⟩. ⟨hal-00494027⟩