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Hello,

If you use 16 bit digitalisation which is an option on the BIORAD it can
happen that you don't see much but you can work with different looking
window and perform all segmentation on the 16 bit images whirout a
compromise.

Bye


----- Original Message -----
From: "Wes Wallace" <[log in to unmask]>
To: <[log in to unmask]>
Sent: Monday, October 28, 2002 7:06 PM
Subject: morphometry of fluorescent images


> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hello Confocalists,
>
> I have a very thorny question about morphometry of fluorescent images.
>
> My question concerns the problem of contrast adjustment and thresholding.
>
> In my experience, the variability of specimen brightness from one
> preparation to another precludes the possibility of setting fixed
> acquisition parameters on the confocal microscope.  For example, I cannot
> acquire all images at the same detector sensitivity, because some
> preparations are brighter than others, and if I did use the same
> detector sensitivity on all, then in some cases the objects being imaged
> would be barely detectable, while in other cases they would be clearly
> detectable.
>
> This leads me to believe that the contrast of each image must be
> normalized after acquisition, so that the objects in each image will be
> represented by a similar range of pixel intensities, and a threshold can
> be applied that is not subjectively chosen for each image.
>
> But is there a way to normalize the contrast of each image that is
> systematic and not biased?
>
> Sincerely,
>
>
> Wes Wallace
> Department of Neuroscience
> Brown University
> Providence, RI 02912 (USA)
> [log in to unmask]
>