Imaging label-free intracellular structures by localisation microscopy.
Localisation microscopy methods allow to realize a light optical resolution far beyond the Abbe-Rayleigh limit of about 200 nm laterally and 600 nm axially. So far, this progress was achieved using labelling with appropriate fluorochromes and fluorescent proteins. Here, we describe for the first time that optical resolution of cellular structures in the λ/10 range (∼50 nm) can be achieved even in label-free cells. This was obtained using Spectral Precision Distance/Position Determination Microscopy (SPDM), a method based on the general principles of localisation microscopy. Besides a substantial resolution improvement of autofluorescent structures, SPDM revealed cellular objects which are not detectable under conventional fluorescence imaging conditions.
Kaufmann
Rainer
R
Applied Optics and Information Processing, Kirchhoff-Institute for Physics, University Heidelberg, Im Neuenheimer Feld 227, D-69120 Heidelberg, Germany.
Müller
Patrick
P
Peptide-Chips and Nucleotide FISH, Kirchhoff-Institute for Physics, University Heidelberg, Im Neuenheimer Feld 227, D-69120 Heidelberg, Germany.
Hausmann
Michael
M
Peptide-Chips and Nucleotide FISH, Kirchhoff-Institute for Physics, University Heidelberg, Im Neuenheimer Feld 227, D-69120 Heidelberg, Germany.
Cremer
Christoph
C
Applied Optics and Information Processing, Kirchhoff-Institute for Physics, University Heidelberg, Im Neuenheimer Feld 227, D-69120 Heidelberg, Germany; Institute for Pharmacy and Molecular Biotechnology, University Heidelberg, Im Neuenheimer Feld 364, D-69120 Heidelberg, Germany; Institute for Molecular Biophysics, The Jackson Laboratory, 600 Main Street, Bar Harbor, ME 04609, USA.
D000818:Animals; D002277:Carcinoma / Q000473:pathology; D045744:Cell Line, Tumor; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D008321:Mammary Glands, Animal / Q000473:pathology; D008856:Microscopy, Fluorescence / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods*
D005456:Fluorescent Dyes