Re: Normalizing intensities

Posted by Gluender on Mar 24, 2007; 7:14pm
URL: http://imagej.273.s1.nabble.com/Normalizing-intensities-tp3699673p3699677.html

Dear commentators!

as far as I can read, the original post didn't mention beads or fluorescence.

With the information provided by the original poster, I should like
to judge the suggested solutions being not lege artis.

>I would argue that in flourescence images the background might
>remain constant even if illumination varies.  Thus for fluor I would
>suggest normalizing to the max value (if your fluor sources don't
>vary much with time), or perhaps the mean intensity of a bright but
>"biologically inert" region of the image.
>
>Bill
>
>-----Original Message-----
>
>From:  Wolfgang Schechinger <[hidden email]>
>Subj:  Re: Normalizing intensities
>Date:  Sat Mar 24, 2007 7:36 am
>Size:  1K
>To:  [hidden email]
>
>Dear Kul,
>
>As differences in light intensities during image recording will
>affect both the background and the peak itensities, I suggest you to
>normalize on the background from all images first, i.e generating
>the normalizing factor for each image based on a reference image in
>order that all images then have the same background. It probably
>makes sense to use the image with the highest background as
>reference as then you won't run into the risk of generating
>overexposed images as the normalization factors will be smaller than
>1. The image with the highest dynamic range also might be a good
>choice if you can determine that easily. This will preserve the full
>dynamics of your image stack.
>
>Can you do a calibration with - eg coloured photobleaching resistant beads?
>
>Best regards,
>
>Wolfgang
>
>-------- Original-Nachricht --------
>Datum: Fri, 23 Mar 2007 14:20:01 -0400
>Von: Kul Karcz <[hidden email]>
>An: [hidden email]
>Betreff: Normalizing intensities
>
>>  Hello,
>>  I have a time series of 2D images. The average intensity of each image
>>  varies slightly from the others, probably because of small changes in the
>>  distance between the microscope's objective and the imaged plane. The end
>>  result is that when you run the time series it flickers.
>>  Can anyone suggest a good way to normalize the intensities, so that the
>>  average intensity throughout the time series is constant?
>>  Thanks,
>>  Kul
>>
>>  Kul Karcz
>>  Corporate Strategic Research
>>  Resource Sciences Laboratory
>>  ExxonMobil Research and Engineering Co.
>
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>
>William A. Mohler
>Dept. Genetics & DevBio
>UConn Health Center
>p: 860-679-1833
>c: 860-985-2719

Best
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                   Herbie

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