Nature Methods
- 5, 539 - 544 (2008)
Published online: 18 May 2008; | doi:10.1038/nmeth.1214
Spherical nanosized focal spot unravels the interior of cellsRoman Schmidt1, Christian A Wurm1, Stefan Jakobs1, Johann Engelhardt2, Alexander Egner1 & Stefan W Hell1, 21
Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, Am Fassberg 11, 37077 Göttingen, Germany. 2
German Cancer Research Center (DKFZ), High-Resolution Optical Microscopy Division, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
Correspondence should be addressed to Alexander Egner aegner@gwdg.de or Stefan W Hell shell@gwdg.de The resolution of any linear imaging system is given by its point spread function (PSF) that quantifies the blur of an object point in the image. The sharper the PSF, the better the resolution is. In standard fluorescence microscopy, however, diffraction dictates a PSF with a cigar-shaped main maximum, called the focal spot, which extends over at least half the wavelength of light ( = 400–700 nm) in the focal plane and > along the optical axis (z). Although concepts have been developed to sharpen the focal spot both laterally and axially, none of them has reached their ultimate goal: a spherical spot that can be arbitrarily downscaled in size. Here we introduce a fluorescence microscope that creates nearly spherical focal spots of 40–45 nm (/16) in diameter. Fully relying on focused light, this lens-based fluorescence nanoscope unravels the interior of cells noninvasively, uniquely dissecting their sub-–sized organelles.
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