I just read a paper and I found that they can also measure fluorescence intensity from the picture.
In an image, each pixels records a number, that is roughly proportional to the amount of photons that reached that particular spot.
From there on, you can follow the logic on how to extract fluorescence intensity from an image...
In an image, each pixels records a number, that is roughly proportional to the amount of photons that reached that particular spot.
From there on, you can follow the logic on how to extract fluorescence intensity from an image...
In addition, this process could depend on certain scale as a reference.
Read this article
Best regards
Article Introduction to the Quantitative Analysis of Two-Dimensional...
Hello,
You can measure fluorescence parameters (mean, median, summary etc) of the total image or its piece and each pixel of the image, of course.
ImageJ could be used for this purpose.
Please use the function of the menu: ANALYZE / MEASURE.
ImageJ from NIH is a free software available from https://imagej.nih.gov/ij/).
They are other commercially available software packages: Volocity, Metamorhp, IPLab, IgorPro, Matlab etc), they are more convenient than ImageJ.
The software allows not only measuring the fluorescence intensity but also some advanced manipulations like subtraction of the background, subtraction one emage from another, summation of 2 or more images, etc. etc. Look at the menu "ANALYZE" in ImageJ.
To correctly compare the images by fluorescence intensity they should be obtained in the same conditions and should be "raw" or processed in the same way.
you should deconvolve the image first, because the objective lense causes a systematic error in the pixel values: The smaller features lose contrast vs. the larger features. So smaller objects have smaller pixel values than they should. Deconvolution mostly fixes this error
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