Leica TCS SP2 Overlay images

[Matthew Pearson-3]

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Hi Everyone,

Does anyone know why in the Leica Confocal Software when you scan multiple fluorophores you select the 'Ovl' button to get an overlay image and then you would press the Single scan button to take an image.  Yet in the Experiment window where the files are saved the overlay image is not saved only the 3 images of each fluorophore (in a triple label).  I have to actually right click the overlay image  and then 'send to experiment, selection raw' in order for it to be saved.  does anyone know if there is a setting to allow the overlay image to be saved without having to save each one manually?  It just seems like a slightly tedious process.

Many thanks,

Matt Pearson.

[Michael Weber-4]

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Matt,

for which type of processing do you need the raw overlay? Overlays are
usually done by the software and not saved (at least not in raw format).

If I remember correctly, you can export the overlay from the experiments
list as ordinary tif or jpg. I am not sure about LCS, but it works in
LAS AF.

Michael


Matthew Pearson wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> Hi Everyone,
>
> Does anyone know why in the Leica Confocal Software when you scan
> multiple fluorophores you select the 'Ovl' button to get an overlay
> image and then you would press the Single scan button to take an image.  
> Yet in the Experiment window where the files are saved the overlay image
> is not saved only the 3 images of each fluorophore (in a triple label).  
> I have to actually right click the overlay image  and then 'send to
> experiment, selection raw' in order for it to be saved.  does anyone
> know if there is a setting to allow the overlay image to be saved
> without having to save each one manually?  It just seems like a slightly
> tedious process.
>
> Many thanks,
>
> Matt Pearson.
>
>
> --
> Imaging Technician
> Cell Biology Division
> Institute of Ophthalmology
> University College London
> EC1V 9EL
> 020 7608 6857
> [hidden email]

[Weis, Michael]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi,

I am a Leica SP2 AOBS user and I feel Leica has got it correct. The data is collected as 8 or 12 bit greyscale in 1-4 channels. This is how the data should be saved, there are many programs that can analyze or merge this data. Overlay works well on the Lieca and if you really want to save it you can. Just remember that trying to extract channel information from an overlay image can be very difficult depending on how many channels there are and the color of the LUT’s.

 

Michael Weis

Electron Microscopy & Digital Imaging

Pacific Agri-Food Research Centre

Agriculture and Agri-Food Canada

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Summerland, BC  V0H 1Z0

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P Before printing think about the environment. / Avant d'imprimer, il faut penser à l'environnement.

 

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Matthew Pearson
Sent: Thursday, April 10, 2008 2:38 AM
To: [hidden email]
Subject: Leica TCS SP2 Overlay images

 

Hi Everyone,

Does anyone know why in the Leica Confocal Software when you scan multiple fluorophores you select the 'Ovl' button to get an overlay image and then you would press the Single scan button to take an image.  Yet in the Experiment window where the files are saved the overlay image is not saved only the 3 images of each fluorophore (in a triple label).  I have to actually right click the overlay image  and then 'send to experiment, selection raw' in order for it to be saved.  does anyone know if there is a setting to allow the overlay image to be saved without having to save each one manually?  It just seems like a slightly tedious process.

Many thanks,

Matt Pearson.

 

-- 

Imaging Technician

Cell Biology Division

Institute of Ophthalmology

University College London

EC1V 9EL

020 7608 6857

[hidden email]