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The pre-edge range background was removed by substracting a Victoreen function and the absorption background was removed using a cubic spline with a normalization range between 70 and 270 eV after the edge.
Fluorescent background was removed using a modified polynomial-fitting method described previously (Beier and Berger2009), making use of the photobleaching lineshape between successive frames as a background fitting parameter.
Raw Cp values were efficiency corrected, and any signal of genomic DNA background was removed using GENEX (http://www.multid.se).se
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LightCycler analysis software was used for quantifications, and background fluorescence was removed using the noise band.
The background noise of the image was removed using the "despeckle" function.
Background or excessively dark haematoxylin staining was removed using the "despeckle" setting and, when required, by superimposing a mask of the haematoxylin channel onto the image.
The false positives generated as a consequence of the oversegmentation tend to be small regions (of less than 200 pixels) representing debris from the background and are removed using region-size-based filtering.
The files were transferred to the AngioSys software (TCS Cellworks), all background and nontubule-like structures were removed using the erode (1 ×) and clean (100 pixels) functions, and the number of pixels representing vessels was counted.
The files were transferred to the AngioSys software (TCS Cellworks), all background and non-tubule-like structures were removed using erode (× 1) and clean (100 pixels) functions, and the number of pixels representing vessels was counted.
The non-reacted Cy-ULS dyes were removed using Agilent KREApure columns to reduce possible background noise for array screening.
The post-edge background was removed by using a derivative method [28,29].
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