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Images were taken with a specimen pixel size of 0.33 nm and a defocus of 2.5 µm.
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Percent collagen was calculated as the ratio of blue-stained to total specimen pixels.
Images were recorded on a 4k Gatan (UK) CCD camera at a magnification of 50,000Χ for negatively stained specimens (pixel size 2.2 Ε; underfocus range: 0.5 1.2 μm) and of 80,000 × for cryo specimens (pixel size 1.4 Å; underfocus range: 1.5 µM–4 μm).
at specimen-level pixel sizes of 0.223 nm or 0.29 nm, respectively, using a dose of ~30 e− Å−2 and a nominal defocus range of −1 to −3 μm.
For example, in fluorescent imaging the signal is often dim with a small number of photons collected from the specimen in each pixel, causing Poisson noise.
This finding was confirmed for the specimen by quantitative pixel-wise comparison of second eigenvector directions and collateral fiber directions assessed on light microscopy image data.
By using a micro CT device, a series of cross-sectional images of the specimen at micrometer-order pixel size are generated by X-rays.
The STOMP macro was used to deliver two-photon excitation light to regions of the specimen corresponding to each pixel in the mask image.
Representative femurs from each treatment group were rescanned using a SCANCO μCT 40 (SCANCO Medical AG) operating at 55 kVp peak energy detection, 6 μm resolution to obtain approximately 2500 cross-sections per specimen in 766 × 763 pixel 16 bit DICOM files.
The average laser power at the specimen was 10 mW, the pixel size was set to 220 nm and a single image consisted of 512 × 256 pixels.
New bone formation within the Inion GTR cylinders was studied by analyzing serial micro-CT images of the specimen at a 12 μm pixel resolution.
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