Re: Ratiometric FRET on Fluoview

Posted by Jean-Pierre CLAMME-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Ratiometric-FRET-on-Fluoview-tp7579865p7579874.html

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Hi Andreas,

The issue is not grouping channels in the sequential box and choosing line
and frame. The issue is changing filter sets quickly. To take the 3 images
DD, DA, AA the filter/light pathway has to be changed between images. DD
and DA  or DD and AA can be taken in the same configuration, but AA and DA
can't. So the only way I can see is to use the Virtual channels and that
is too slow line by Line.

Best

JP


 


Confocal Microscopy List <[hidden email]> wrote on
03/05/2013 07:01:06 AM:

> Andreas Bruckbauer <[hidden email]>
> Sent by: Confocal Microscopy List <[hidden email]>
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> 03/05/2013 07:05 AM
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> Confocal Microscopy List <[hidden email]>
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> Re: Ratiometric FRET on Fluoview
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> I have not used it for FRET, but we have a sequential tick box at
> the bottom of the window with the channel parameters. When ticked we
> can select "frame by frame" or "line by line" aquisition and how the
> channels will be grouped together.

> best wishes

> Andreas

>
> -----Original Message-----
> From: Jean-Pierre CLAMME <[hidden email]>
> To: CONFOCALMICROSCOPY <[hidden email]>
> Sent: Mon, 4 Mar 2013 22:44
> Subject: Ratiometric FRET on Fluoview

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> Hello,

> I saw a paper about Ratiometric FRET (Roszik, Cytometer 2009) mentioning
> line by line acquisition of the 3 images (IDA, IDD, IAA). The authors
> mentioned the use of a LSM 510.

> I don't know the LSM510, but I have a fluoview 1000 and the only way I
can
> see how to do that is using the "virual Channels" . However that would
be
> image by image and not line by line.

> Does anyone has done ratiometric FRET on the fluoview and what method
did
> you use ?

> Thank you and Best regards,

> JP

>
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> Jean-Pierre CLAMME, PhD
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