Picard 4D stage with high NA detection lens

Hi all,

Just wondered who is using the Picard 4D stage on their SPIM with a high NA detection lens? The z axis has a minimum step of 1.5µm which is a lot higher than it should be for Nyquist sampling in z with high NA objectives. This then becomes the limiting factor in z resolution. Having said that if you did use Nyquist you would extend the imaging time and capture significantly more data given the size of light sheet samples.

Thanks,

Matt

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Hi @Matt_Pearson1,

as far as I know, you can run the stage with a continuous speed between defined positions. If you calibrate that speed with the light exposure you should be able to acquire slice distances smaller than 1.5 micron. However I could also imagine some motion blur from this “continous” acquisition. How much freedom do you have in programming your scope? I guess you’re using micromanager?

Cheers,
Robert

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Hi Robert,

Yes I see what you’re saying, capture the images as the stage is moving between these defined points. I guess you would have some blur also if you wanted to capture multiple wavelengths as sequential exposures the stage will have moved a bit by the time it captures channel 2. Yes we’re using Micro-Manager so not a closed box if you know what you’re doing.

Thanks,

Matt

@Matt_Pearson1 Just as an aside, in case you weren’t aware: there is also a new forum for discussing acquisition and microscopy hardware questions: μForum. Questions about the hardware aspects (OpenSPIM or otherwise) would be well-received there. That said, software-oriented questions about OpenSPIM, μManager, etc., are perhaps more appropriate here. For these bioimaging projects straddling software and hardware, the line is a bit blurry.