I am a research student and taking initiative to do some quantitative histological stain analysis with Fiji-imageJ. I have performed histological stain- masson trichrome with aniline blue (that demonstrates) collagen fibres as blue. My aim is to determine the density of the collagen present in a defined ROI. I have been able to set scale, RGB colour space conversion, colour image image deconvolution using ‘masson trichrome’ vector. Now my challenges are as follows:
- Is it necessary to colour ‘Threshold’ for quantification as there is already colour separation during image deconvolution?
2.In selecting a ROI to measure area, IntDen and RawInDen, am I to say IntDen=colour intensity per that selected area=density of fibres in that selected area? EG 1 Area=17667.98 Mean= 130.95 StdDev= 61.53 Min7 Max=245 IntDen= 2313621.99 RawIntDen= 80539752.00
- How can I deduce the density of collagen fibres from the data obtained if my scale set was at 5.87pixels/um.
Please do help as I am a beginner to imageJ analysis but finding the software interesting and very useful.
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