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Color thresholds were set to identify positive labeling above background and colocalization analyses were performed in ImageJ in user-defined regions of interest (ROI) using the coloc2 plugin and a Costes P-value of 1, indicating that colocalization is not random.
Then, using MetaMorph® Microscopy Automation & Image Analysis Software (Molecular Devices, LLC, Sunnyvale, CA, USA), color thresholds were defined for the entire tumor area and the specific antibody bound tissue only.
Besides, the combination of different color thresholds may achieve sufficient detection rate on embedded system by using a small amount of hardware resources (Liu and Shi, [2011]), but such system can only work under adequate illumination and tedious calibration is still unavoidable because of the lack of adaptive mechanisms.
For assessing cytoplasmic staining, the area positive stain was calculated by applying color thresholds to detect positive brown pixels.
Individual cells were discriminated by incorporating the shape and size thresholds, providing, together with the color thresholds, actual cell counts.
For assessing nuclear staining, positive DAB staining was calculated by applying two color thresholds with one recognizing blue background (haematoxylin stained) cells and another recognizing brown positive cells and blue, non-positive cells (total cell number).
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Mr. Meyerowitz had crossed the color threshold never to turn back.
As an arbitrary setting to measure the stained area, a color threshold ranging from 160 to 255 in brightness was selected.
In detection step, the transcendental color threshold method is used for initial filterings, meanwhile, the prior knowledge is performed to scan the scene in order to quickly establish candidate regions.
After described the H-S plan characteristics, next we can determine an adaptive skin color threshold by using the TCH algorithm, which can be discussed as in the following cases.
Although it also suffers from the non-uniformity problem where the same distance between two color points within the color space may be perceptually quite different in different parts of the space, within a certain color threshold it is still definable in terms of color consistency.
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com