Posted by
Joel Sheffield on
Feb 28, 2020; 5:35pm
URL: http://imagej.273.s1.nabble.com/HELP-segmentation-and-quantification-of-DIC-images-tp5022977p5022990.html
The image arises from a DIC optical system. DIC images of spherical
objects show the shadow like appearance that you see in each of the
particles. It is often very difficult to get a uniform background in these
images as well. So, the problem that you face is twofold. 1. smooth the
background, and 2. measure the particles, given that they are distorted by
the shadows. ImageJ has some routines that might be helpful in this. As
has already been suggested, you can use the FFT>bandpass filter to smooth
the background, since the background variation can be thought of as a long
wavelength curve of intensity, and so you are removing the low frequency
component of the image. Measuring the particles is trickier. However, it
seems to me that you can use the Process>filters>variance routine to make
the shadows equivalent from both sides. The problem with this is that it
elongates the image. On the other hand, since you would now have a
population of ellipses, you could threshold them and then use the Particle
Analyzer to look for the short axis of each ellipse. These are just
ideas, since I haven't actually tried them out.
Best of luck,
Joel
Joel B. Sheffield, Ph.D
Department of Biology
Temple University
Philadelphia, PA 19122
Voice: 215 204 8839
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On Thu, Feb 27, 2020 at 7:10 AM Hui Zhang <
[hidden email]> wrote:
> <
http://imagej.1557.x6.nabble.com/file/t382540/DIC-droplets.png> Hi
>
> I am a phD student in Germany. recently I am focusing on the phase
> separation property of a protein.
> As tested the GFP tag can enhance the ability of the protein to phase
> sepatate, thus forming bigger droplets.
> So I can only purified and utilize the untaged protein for assays and
> generate a series of two dimentional DIC images with round shapes.
> now the problem is that I need to quantify the number, size and roundess
> (aspect ratio), which requires a good program to segmentate.
> I attached an image also in case you want to have a look. you can see that
> the background is not even, maybe it is not qualified, Do I need to
> generate
> new images?
>
> does someone of you know how to process or have some good program,
> software,
> or plug-in that I can apply.
>
> Best wishes
> Hui Zhang
>
>
>
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