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. 2011 Jun 6;50(24):5581-3.
doi: 10.1002/anie.201100371. Epub 2011 May 6.

Fluorescence nanoscopy of single DNA molecules by using stimulated emission depletion (STED)

F Persson  1 , P Bingen, T Staudt, J Engelhardt, J O Tegenfeldt, Stefan W Hell
Affiliations
Free PMC article

Fluorescence nanoscopy of single DNA molecules by using stimulated emission depletion (STED)

F Persson et al. Angew Chem Int Ed Engl. .
Free PMC article
No abstract available

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Figures

Figure 1
Figure 1
a) Confocal image of YOYO stained λ-DNA (basepair:dye 5:1). b) The corresponding STED image taken with λSTED=568 nm (raw data). The STED image was acquired before the confocal counterpart. Scale bars: 1 μm. c) Average of three line profiles from the STED (solid red line) and confocal (dotted black line) images. Line profiles extracted along the white lines in (a) and (b). The three distinct peaks belonging to different DNA molecules are only resolved by STED.
Figure 2
Figure 2
Typical raw STED images of YOYO stained λ-DNA (basepair:dye 5:1) using a) λSTED=568 nm and b) λSTED=647 nm. Scale bars in (a) and (b): 1 μm. c) Graph showing the average of 11 line profiles of a single DNA strand, with fits, for standard epifluorescence (dotted light gray line), confocal (dashed black line), and STED nanoscopy with λSTED=647 nm (dash-dotted blue line) and 568 nm (solid red line); the corresponding full width at half maximum (FWHM) were found to be (300±11) nm (Gaussian), (238±5) nm (Gaussian), (62±2) nm (Lorentzian), and (42±3) nm (Lorentzian), respectively. The error bars correspond to one standard deviation. d)–g) Examples of DNA segments with bends and kinks visible (indicated by white arrows) in a STED image using e) λSTED=647 nm and g) λSTED=568 nm but not resolvable in the corresponding confocal images (d) and (f). Scale bars in (d)–(g): 500 nm.
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References

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