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Re: Thresholding help

Posted by Michael Schmid on Oct 30, 2008; 12:04pm
URL: http://imagej.273.s1.nabble.com/Thresholding-help-tp3694620p3694624.html

Hi Justin,

the ImageJ built-in "subtract background" command should work well  
with your type of image.

It will be very difficult to extract the weakly visible sides of the  
particles. For a rough size estimate of the circular particles, a way  
out might be checking the width of the rectangular bounding box and  
Feret's diameter - if these sizes agree within reasonable limits,  
they shuld be ok. For the remaining particles you have to do it  
manually.

Here is a macro for the first steps:
   run("32-bit");
   run("Smooth");
   run("Subtract Background...", "rolling=100 light sliding");
   setThreshold(-9999, -10);
   run("Set Measurements...", "  bounding feret's redirect=None  
decimal=3");
   run("Analyze Particles...", "size=10-Infinity  
circularity=0.00-1.00 show=[Nothing] display include record");

If this is not enough, you need either better illumination or a  
plugin that fits full circles into the arcs (I am not aware of any,  
you might have to write it yourself).

Michael
________________________________________________________________
Michael Schmid                    email: [hidden email]
Institut fuer Allgemeine Physik, Technische Universitaet Wien
Wiedner Hauptstr. 8-10/134, A 1040 Wien, Austria
Tel. +43 1 58801-13452 or -13453, Fax +43 1 58801 13499
________________________________________________________________

On 29 Oct 2008, at 22:02, Michael Doube wrote:

> Justin:
>
> How about subtracting the background gradient from the image to  
> improve contrast?
>
> Just take a 'background' image without any droplets and use Process-
> >Image Calculator, subtracting your background from the droplets
> +background image.  I had a quick go on your example and greatly  
> improved the ability to discriminate the droplet contours.
>
> Mike
>
> Fabrice Senger wrote:
>> Quoting Justin Walker <[hidden email]>:
>>> I have a series of images of liquid droplets, which have very poor
>>> contrast with the surrounding phase, due to the similarities in  
>>> their
>>> index of refraction.  I get enough contrast in my images to  
>>> clearly see
>>> the droplets, however, I can't pick them out with the threshold  
>>> because
>>> there is a slight gradient in the brightness of my lighting.  
>>> Link to a
>>> sample image is below:
>>>
>>> http://www.chejrw.com/forum/test.tif
>>>
>>> Does anyone know of a way I can manipulate this image to be able  
>>> to use
>>> the 'measure particles' plugin to get the droplet sizes?  I can  
>>> clearly
>>> see the droplets, so surely there must be a way to make the computer
>>> see them.
>>>
>>> Thanks everybody
>>>
>>> - Justin Walker
>> As far I can see in your images, you can see the "contour" of the  
>> droplets.
>> If you invert your image, this "contour" should appear bright in a  
>> dark background.
>> This should help.
>> Fabrice.