FRAP: measure intensity of moving foci problem. Is there a moving ROI application in ImageJ

[MartijndeG]

Hello everybody,

I'm working with ImageJ for some time and I need a certain function/macro/plugin for my research.

I bleach certain parts of foci with "FRAP on the Fly" and when I analyse the image stack (.stk MetaMorph), I form a ROI over the bleached region and measure the intensity during the recovery of the fluorescence signal.

The problem is that these foci are moving, not much, but in the period of measurement they migrate their own size in distance, so measuring this bleached region with a ROI results in lost of signal due the migration out of the ROI.

Is there a solution for this problem, I can understand this happens with more FRAP experiments?
I thought myself there must be an application to let the ROI move over a certain predefined distance, to cover the bleached region.

many thanks in advance!

[Johannes-P. Koch]

I used a plugin called FRAP-Profiler which is part of the MBF collection.
I encountered a similar problem: I bleached part of organellar substructures
and of course this structure moved. I do not know whether this is of any
help for you, I used a registration plugin to "fix" the moving structure
rather than let the ROI move. This was possible because i) I observed no
recovery and ii) the overall structure stayed the same throughout the
observation period.

Of course, you could move the ROI manually and measure all the time
separately....this might be the easiest solution since if you wanted to
apply a moving ROI you would have to tell it how fast and where to move....

Best,
Johannes


----- Original Message -----
From: "MartijndeG" <[hidden email]>
To: <[hidden email]>
Sent: Thursday, October 29, 2009 3:16 PM
Subject: FRAP: measure intensity of moving foci problem. Is there a moving
ROI application in ImageJ

> Hello everybody,
>
> I'm working with ImageJ for some time and I need a certain
> function/macro/plugin for my research.
>
> I bleach certain parts of foci with "FRAP on the Fly" and when I analyse
> the
> image stack (.stk MetaMorph), I form a ROI over the bleached region and
> measure the intensity during the recovery of the fluorescence signal.
>
> The problem is that these foci are moving, not much, but in the period of
> measurement they migrate their own size in distance, so measuring this
> bleached region with a ROI results in lost of signal due the migration out
> of the ROI.
>
> Is there a solution for this problem, I can understand this happens with
> more FRAP experiments?
> I thought myself there must be an application to let the ROI move over a
> certain predefined distance, to cover the bleached region.
>
> many thanks in advance!
>
>
> --
> View this message in context:
> http://n2.nabble.com/FRAP-measure-intensity-of-moving-foci-problem-Is-there-a-moving-ROI-application-in-ImageJ-tp3912741p3912741.html
> Sent from the ImageJ mailing list archive at Nabble.com.
>
>

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