I have to quantify the fluorescence of an image by comparing the pixels of fluorescent and normal image which can be done by ImageJ software which is freely available. However I could not do the fluorescence determination of an image using this software. Can anyone make any suggestions in this regard?
In ImageJ software, pixel counts are used to determine the differences in intensity of flourescence. fluorescence percentage determined by dividing the no. of flouerescent pixels of a nematode(an organism) by total pixel area. a threshold is being set to obtain the no. of flourescent pixels for each image. threshold is set in control non-fluorescing images. but how the threshold value is set and how it is proceeded further is unclear to me.
the flowchart showing the different functions of imageJ software in chronological order will be helpful.
i could proceed up to:
file-open-image-adjust-threshold. in threshold 2 bars are there showing Default & Over/Under. after that i am lost. means how to set threshold and then how to count the florescent pixels in other images.
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