Re: intensity measurements
Posted by
Julio Vazquez on
Jun 01, 2009; 4:16pm
URL: http://confocal-microscopy-list.275.s1.nabble.com/intensity-measurements-tp3005594p3007096.html
Hi Vaishali,
The method I find the most reproducible and satisfactory is to separate the color channels and then perform intensity measurements in the channel that best separates your specific stain. Basically, you analyze your image as if the DAB staining were a fluorescence (intensity) channel. You can find a description and example in Pham et al., Diagnostic Pathology 2007, 2:8, Quantitative image analysis of immmunohistochemical stain using a CYMK color model.
In this example, they convert their images to CYMK and analyze the yellow channel, but basically you want to pick a channel that maximizes the intensity of your strain and minimizes the intensity of the background. You could do that with RGB, or other color separation methods. This approach is easy to implement with imagej and most other image analysis software. You may have to invert your image so that your intensity measurements are positive (that is stain higher than background).
The other approach is to try to separate colors using a color thresholding method, for example with the "threshold color" plugin of imageJ. I find this more difficult to work with and more unreliable, generally.
good luck,
Julio.
--
Julio Vazquez
Fred Hutchinson Cancer Research Center
Seattle, WA
== On Jun 1, 2009, at 3:51 AM, vaishali kailaje wrote:
hello friends,
We are interested in measuring the intensity in immnohistochemical slides (DAB stained).
Could anybody suggest some software or a method to quantitate DAB intensity in tissue sections.
Secondly,can these measurements be made using metomorph, ImageJ or Axiovision as these are the softwares available with us.
Thanks
Vaishali Kailaje
Scientific Assistant,
Tata Memorial Centre,
Advanced Centre for Treatment Reasearch Education in Cancer
Kahrghar,
Navi Mumbai,
India.