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Hi Emmanuel,

If you stop the disc, while imaging, you will see the pinholes. 

Adjust the camera, sharpest image/intensity while looking at this image. 

Hope this helps. 


Best,

Gary

> On Apr 16, 2024, at 5:23 PM, Emmanuel Levy <[log in to unmask]> wrote:
> 
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> Post images on http://www.imgur.com and include the link in your posting.
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> 
> Dear Zdenek and Cătălin,
> 
> Thank you for your replies! Indeed, the "distance" of the camera relative
> to the camera-port lens sounds like an intuitive parameter to adjust.
> 
> So the question is, have you adjusted that distance before and achieved a
> 1-2 pixel zoom in/out in this manner? I assume that there should also be an
> optimal distance for proper focus
> 
> The picture I attached was imaged with beads. We do a lot of
> co-localization experiments, so it would be good if we managed to tune the
> hardware flawlessly.
> 
> Thank you for sharing your experience.
> Best wishes,
> 
> Emmanuel
> 
> 
>> On Mon, 15 Apr 2024 at 17:31, Cătălin Pavel <[log in to unmask]> wrote:
>> 
>> *****
>> To join or leave the confocal microscopy listserv or to change your email
>> address, go to:
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>> *****
>> 
>> Hi Emmanuel,
>> It looks that one of the camera is closer to the microscope than the
>> other. Try to check that all the connections are flush and tight.
>> 
>> Catalin
>> 
>>> On Apr 15, 2024, at 09:30, Emmanuel Levy <[log in to unmask]>
>> wrote:
>>> 
>>> *****
>>> To join or leave the confocal microscopy listserv or to change your
>> email address, go to:
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>>> Post images on http://www.imgur.com and include the link in your
>> posting.
>>> *****
>>> 
>>> Dear All,
>>> 
>>> We have a W1 spinning disk with dual cameras. We recently had them
>> aligned,
>>> and upon close inspection, we see that the GFP channel image is about 1
>> to
>>> 2 pixels larger than the RFP image in every corner (we used a 60x
>>> objective, and the cameras are primeBSI express with 6.4um pixels, so the
>>> image is off by 100-200nm in every corner). you can download the two
>> images
>>> as a composite here if you are curious:
>>> 
>> https://drive.google.com/file/d/1TWIxwiZF6uf3FspyA96AX6r092PB_102/view?usp=sharing
>>> 
>>> What would be the best way to solve this issue? Shouldn't the standard
>>> camera mounts enable us to correct this?
>>> 
>>> Thanks for your help and comments,
>>> Best wishes,
>>> 
>>> Emmanuel
>>