http://imagej.273.s1.nabble.com/Help-with-quantifying-pictures-after-channel-splitting-tp5011445p5011531.html
She is using the blue channel correctly. In the brightfield image red is
presence of red. The dye does not add additional red light but absorbs away
blue and/or green. 😃
На 08.02.2015 04:53 "G. Esteban Fernandez" <
> Hi Stephanie,
>
> For this kind of thing the Colour Deconvolution plugin usually works well
> for me (if staining is consistent from image to image). The plugin would
> separate your color images into two images corresponding to the two
> histological stains you used.
>
> In addition to Colour Deconvolution I'd apply a Process > Subtract
> Background step (right now I don't remember if background subtraction works
> better before or after color deconvolution).
>
> You could also try the Auto Local Thresholding plugin instead of the global
> thresholding you described.
>
> In short, my workflow is generally this:
>
> 1. Colour Deconvolution (comes with Fiji ImageJ)
> 2. Process > Subtract Background
> 3. Auto Local Threshold (also in Fiji)
>
> -Esteban
> On Feb 4, 2015 6:58 AM, "Stephanie Klett" <
[hidden email]>
> wrote:
>
> > Dear Image J discussion group,
> > I have a question about how I can quantify my digital microscopic images
> > (.jpg) In my case I done a immunological staining where red is the colour
> > of
> > interest. Therefore, I split the channels and use the blue labelled
> picture
> > and apply the function Threshold... (under Image-->Adjust-->). Then I
> get a
> > histogram and an automatically adjustment of the colour sensibility (?).
> My
> > question is what should I keep in mind when I want to compare several
> > images. Do I have to adjust always the same sensibility (from the rage
> of 0
> > to 255) or do I choose the automatically adjustment? My problem is, that
> > sometimes the automatically adjustment represent the quantification
> better
> > than the sensibility rate that I decided to use and other way around.
> Which
> > way is more unbiased or do I have to do a completely other procedure?
> >
> > Here I display screen shots where I document my way and may problem more
> > exemplified.
> >
> > Fig. 1 Opened a file and split the channels
> >
> > Fig. 2 Apply the function ³Threshold...² on the image labelled "blue³
> >
> > Fig. 3 Display the automatically adjustment of 228
> >
> > Fig. 4 Corrected the threshold to 180
> >
> > Fig. 5 A new image, display the automatically threshold of 222
> >
> > Fig. 6 Corrected the threshold to the undoing rate of 180
> >
> >
> > I¹m very happy if you can help me and looking forward to your answer.
> >
> > Thank you for your attention
> >
> > Best regards
> > Stephanie
> >
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http://imagej.nih.gov/ij/list.html> >
> >
>
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