Stack bates resolution easily

Aug 6th, 2022
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How to stack bates resolution

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Fluorescence microscopy is a valuable toolbox to study cellular structures and dynamics spanning scales from the single molecule to the live animal. The spatial resolution that can be achieved with any light-based microscopy is however limited to about 200 nm in the imaging plane and 500 nm along the optical axis.
By exciting the fluorophores or detecting the signal through the nonpropagating light near the fluorophore, high-resolution information be retained. Near-field scanning optical microscopy (NSOM) acquires an image by scanning a sharp probe tip across the sample, typically providing a resolution of 2050 nm (911).
Resolution of the light-sheet microscope 26 m, depending on the field of view) in the case of the commonly used long working distance objectives with large depth of focus. With a 10x NA 0.3 objective the axial resolution of a light-sheet microscope is better than that of a laser scanning confocal.
The best resolution for an optical microscope is about 0.2 microns = 200 nm.
STORM can resolve structures with sub-diffraction-limit resolution. (a) STORM cleanly resolves two switches separated by a contour length of 46 nm on double-stranded DNA. The STORM images show two clearly-separated clusters of measured switch positions (crosses), each corresponding to a single switch.
In fact, the physicist Ernst Abbe had formulated in 1873 that the resolution of light microscopes is limited to half the wavelength of light, which is about 200 nanometers. More than 100 years later, this Abbe limit is still valid.
Conventional microscopy techniques, namely the confocal microscope or deconvolution processes, are resolution limited, ~250 nm, by the diffraction properties of light as developed by Ernst Abbe in 1873.

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