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It takes advantage of the high sensitivity of polarimetry for optically anisotropic structures, even below the optical resolution, such as for the 30 nm-diameter stromal collagen fibrils.
It shows that the fibril diameter is always below the optical resolution, in agreement with TEM images.
SHG microscopy is then sensitive to fibrils with diameters below the optical resolution (360 nm in our set-up in the lateral direction).
Moreover, we showed that we could focus on the growth of a single isolated collagen fibril because SHG microscopy is sensitive to well-organized fibrils with diameter below the optical resolution.
Isolated single molecules or complexes with a size below the optical resolution (a few hundred nanometers) appear as spots whose shape is dictated by the point-spread function (PSF) of the microscope.
Polarization resolved techniques are indeed highly sensitive to the sub-micrometer distribution of anisotropic elementary components within a sample, much below the optical resolution, as routinely exploited in ellipsometry measurements.
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The resolution performance of a sensor with submicron pixels exceeds the optical resolution limit.
In addition, we use Airyscan super-resolution microscopy (~140 nm-resolution) to bring the optical resolution beyond the diffraction limit of light16,17,18.
The optical resolution limit for these images was 0.48 µm in the XY direction.
Contrary to many classical microscopy methods, the structural resolution in localization microscopy is potentially much lower than the optical resolution.
The specimen was submerged in water to improve the optical resolution.
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