Huge delay between images when using Zen 3.1 and Zeiss Fast Dual Filter wheel

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Bradley Dyer Bradley Dyer
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Huge delay between images when using Zen 3.1 and Zeiss Fast Dual Filter wheel

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Dear Confocal List Serve Colleagues,

I have been attempting to run time lapse acquisitions using Zeiss' Zen 3.1 software for a number of weeks now with little success.  I would like to be able to acquire in at least 2 channels, using an Axio Observer 7 fitted with the fast dual filter wheel assembly, with as little time delay between cycles as possible.  The filter wheel is supposed to be able to change positions in 50ms.

In reality the total overhead has been between 250 and 450ms per image acquired meaning short, 100 cycle experiments are taking minutes to complete.

If anyone has any experience with this set up then any help in making these experiments run faster would be greatly appreciated!

Thank you in advance,

Microscopy team at Ward Ober Lab
Csúcs  Gábor-3 Csúcs Gábor-3
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Re: Huge delay between images when using Zen 3.1 and Zeiss Fast Dual Filter wheel

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Dear Bradley,

Unfortunately, I do not have a solution for running your experiments faster unless you are ready to change a component (hardware or software) of your imaging system. Others might have better suggestions but I would see 3 possibilities:

1) Try a different software (e.g. Metamorph or MicroManager).  According our experience the various software are good at controlling various components...
2) Try to not use the filter wheel, rather use an LED light source and a multi-band dichroic/emission filter.
3) Use a hardware solution like the one offered by InscopeR that separates the control of the various hardware components from the software and hence enables you to use your hardware at its full speed. Unfortunately, also the InscopeR solution requires that you change your imaging software.

Greetings Gabor

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Bradley Dyer
Sent: Wednesday, 26 May 2021 17:19
To: [hidden email]
Subject: Huge delay between images when using Zen 3.1 and Zeiss Fast Dual Filter wheel

*****
To join, leave or search the confocal microscopy listserv, go to:
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Dear Confocal List Serve Colleagues,

I have been attempting to run time lapse acquisitions using Zeiss' Zen 3.1 software for a number of weeks now with little success.  I would like to be able to acquire in at least 2 channels, using an Axio Observer 7 fitted with the fast dual filter wheel assembly, with as little time delay between cycles as possible.  The filter wheel is supposed to be able to change positions in 50ms.

In reality the total overhead has been between 250 and 450ms per image acquired meaning short, 100 cycle experiments are taking minutes to complete.

If anyone has any experience with this set up then any help in making these experiments run faster would be greatly appreciated!

Thank you in advance,

Microscopy team at Ward Ober Lab
George McNamara George McNamara
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Re: Huge delay between images when using Zen 3.1 and Zeiss Fast Dual Filter wheel

In reply to this post by Bradley Dyer
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Claire Brown has published that going through USB interface can cause
massive delays. If you are connected through a USB3 port, it is possible
that the connection is not operating at full speed. If you have parallel
port (direct, not USB!) or TTL, try those. Also previous person's post
suggestion trying alternative software (hmmm ... was a previous version
of Zen fast?).

happy 2021,

George

On 5/26/2021 11:19 AM, Bradley Dyer wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Confocal List Serve Colleagues,
>
> I have been attempting to run time lapse acquisitions using Zeiss' Zen 3.1 software for a number of weeks now with little success.  I would like to be able to acquire in at least 2 channels, using an Axio Observer 7 fitted with the fast dual filter wheel assembly, with as little time delay between cycles as possible.  The filter wheel is supposed to be able to change positions in 50ms.
>
> In reality the total overhead has been between 250 and 450ms per image acquired meaning short, 100 cycle experiments are taking minutes to complete.
>
> If anyone has any experience with this set up then any help in making these experiments run faster would be greatly appreciated!
>
> Thank you in advance,
>
> Microscopy team at Ward Ober Lab
Armstrong, Brian Armstrong, Brian
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Re: Huge delay between images when using Zen 3.1 and Zeiss Fast Dual Filter wheel

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Hi, we had a similar problem. We were able to sinc the camera to the Colibri via the trigger board. If you have the fast components: trigger board, trigger cable and signal distribution box, as well as the time lapse module in the software then you should able to have a Zeiss rep walk you through the set-up. It was configured incorrectly after the install and an application specialist from Zeiss corrected it for us.
Hope this helps.

Brian Armstrong PhD
Director, Light Microscopy Core
Research Professor
Department of Shared Resources
Beckman Research Institute, City of Hope

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From: Confocal Microscopy List <[hidden email]> On Behalf Of Bradley Dyer
Sent: Wednesday, May 26, 2021 8:19 AM
To: [hidden email]
Subject: Huge delay between images when using Zen 3.1 and Zeiss Fast Dual Filter wheel

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Dear Confocal List Serve Colleagues,

I have been attempting to run time lapse acquisitions using Zeiss' Zen 3.1 software for a number of weeks now with little success.  I would like to be able to acquire in at least 2 channels, using an Axio Observer 7 fitted with the fast dual filter wheel assembly, with as little time delay between cycles as possible.  The filter wheel is supposed to be able to change positions in 50ms.

In reality the total overhead has been between 250 and 450ms per image acquired meaning short, 100 cycle experiments are taking minutes to complete.

If anyone has any experience with this set up then any help in making these experiments run faster would be greatly appreciated!

Thank you in advance,

Microscopy team at Ward Ober Lab

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