i am new to using Imaris and i have a few question regarding thresholding during image processing. When is using the background subtraction in Imaris relevant for the analysis of the image? Do you enter the diameter of the smallest object or the largest object into the threshold value?
Hi,
Thresholding is very “violent” technic to obtain a binary image whatever the quality of the document that must be analyzed. Background subtraction is useful when the background is easy to be quantified everywhere in the document, and this is not always possible. The best approach is to manipulate directly the available filters and specifically those that calculate locally the areas of interest in the image and/or a organized cascade of them. This requires a good knowledge of image mathematics and the use of images convolutions, whatever the advertising arguments that can be red… Good luck!
Christian
HI
I also have similar question
We set up the experiment having test and negative control
My concern is how to take negative control into account to decide thresholding subtraction from test image.
I mean if I have control image with no florescent dye and getting some autofluroscence, I tried to adjust the parameters to get null background and out same parameters to analyse the test image.
Now for this approach again question comes as every image is different in confocal slightly in terms of section of z stack and all, what we see there is normalised grey scale value with reference to image.
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