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Comment
. 2000 Aug 1;97(16):8747-9.
doi: 10.1073/pnas.97.16.8747.

Shattering the diffraction limit of light: a revolution in fluorescence microscopy?

S Weiss  1
Affiliations
Comment

Shattering the diffraction limit of light: a revolution in fluorescence microscopy?

S Weiss. Proc Natl Acad Sci U S A. .
No abstract available

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Figures

Figure 1
Figure 1
A nonlinear relationship between the fluorescence emission (F) and the excitation light (Iex) “shrinks” the emission PSF. An excitation “input” PSF (blue, Left) will be converted into an emission “output” PSF (green, Right) via a “nonlinear fluorescence box” (Center). The Jablonski diagram at the top right of the “fluorescence box” describes an arbitrary nonlinear excitation process (although only one wavy arrow is shown for Iex, it can describe a multiphoton process). The output PSF is normalized and compared with the input PSF.
Figure 2
Figure 2
PSFE by STED. (a Left) A Jablonski diagram showing the ground (S0) and the first excited singlet (S1) states of a fluorophore with the corresponding vibrational manifolds. The fluorophore acts as an effective four-level system because the vibrational relaxations of S1 and S0 are fast compared with the fluorescence lifetime. A short pulse (Iex, 200 fs) excites the fluorophore into a high vibrational state of S1. The slow STED pulse (STED, 40 ps) stimulates emission from the lowest S1 vibrational state into a high vibrational state of S0. The stimulated emission depletes S1 and quenches the fluorescence. (Right) The equivalent wavelength diagram showing the absorption curve (dotted black line), the emission curve (solid black line), and the positions of Iex, STED and the filter (F). The filter blocks the two lasers and passes the signal (hatched green). (b) The combination of the excitation PSF (blue) with the STED engineered intensity distribution (red) acts as an input to the PSFE by STED “nonlinear box.” The output PSF (green) is shrunk.
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