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November 2007

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From:
Glyn Nelson <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Fri, 9 Nov 2007 10:45:55 -0500
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Xuejun is right, this works well to produce a pseudo colour transmission 
image.  We have used it to obtain detailed high resolution images of 
histological sections (our normal widefield not having as nice 
objectives).  The normal tilescan does limit you to 4096 x 4096 pixels I 
think, but you can overcome this by using the tilescan feature in the 
multitime macro if you have a motorised stage.

Glyn.

On Thu, 8 Nov 2007 09:47:55 -0700, Xuejun Sun <[log in to unmask]> wrote:
>Hi, Alice,
>
>We did not use the Zeiss camera. However, we had a similar need long time
>ago as some one tried to correlate histology with immunofluorescence. 
Here
>was how we got by (without a camera): use your red, green and blue laser
>lines and set up a sequential scanning configuration with your 
transmitted
>detector. You can get a pseudo-color image which might need some tweaking
>(mostly gamma) by the end. Most importantly, you can just use the tilscan
>function of your LSM and get your color tiled images. If you calibrate 
the
>stage, you can get a reasonable tiled image out of it. But I think Zeiss
>software limits the final frame size (in term of number of pixels).
>
>It has been a long time we did this and I might forget some details. 
Initial
>setting up (gain/offset of the detector) was a bit pain in term of 
getting
>correct colors. But once set, you can reuse it and tweak the final image 
in
>photoshop. Additionally, If I recall correctly, one critical thing is to
>remove DIC optics from the optical path or you might get a gradient in 
your
>tiled image.
>
>Good luck,
>
>Xuejun
>
>
>Xue-jun Sun, Ph.D.
>Dept. Exp. Oncology
>Cross Cancer Institute
>11560 University Ave,
>Edmonton Alberta T6G 1Z2
>Canada
>Phone: (780) 432-8898
>Fax:     (780) 432-8425
>
>-----Original Message-----
>From: Confocal Microscopy List [mailto:[log in to unmask]] On
>Behalf Of Alice L. Givan
>Sent: Wednesday, November 07, 2007 3:26 PM
>To: [log in to unmask]
>Subject: CCD camera
>We would like to add a color CCD camera to our Zeiss LSM-510 META 
confocal
>to capture color tiled wide field images.  We wondered if other users 
have
>already done this?  What cameras did you add and how well does the LSM 
510
>software control the image acquisition with the camera you are using?  We
>are particularly interested in tiled images.
>
>We have a PixeLINK model PL-A662 color CCD camera on a separate stereo
>scope.  We are told that this camera would not work with the LSM-510.  
Does
>anyone have any contrary information about this?
>
>Thanks for your  help.
>
>Alice
>
>Alice L. Givan
>Englert Cell Analysis Laboratory
>of the Norris Cotton Cancer Center
>Dartmouth Medical School
>Lebanon, NH 03756 USA
>tel 603-650-7661
>fax 603-650-6130
>[log in to unmask]
>www.dartmouth.edu/~celllab

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