Re: [micro-manager-general] Questions on Hamamatsu Flash4

[julou <tho...@no...>]

Hello,

Thanks Mark for your answer.
Sabrina, I unfortunately have no experience with .dcimg files… By the way, it’d have been nicer to have started a dedicated thread since this is an independent question.

> Performing acquisitions at precise intervals that are not dictated by  the camera (based on exposure and other settings) requires either (a) a camera that supports adjustable intervals when in streaming mode, with µManager support for such functionality, or (b) external triggering of the camera using an appropriate device. Unfortunately (a) is not available for Hamamatsu cameras in µManager, and neither (a) nor (b) is supported in the µManager GUI ((b) is on our eventual roadmap) and therefore would require scripting. 

> > Does this mean that the only way for us to speed up z-stacks acquisition would be to use a triggerable z-stage? 
> 
> Yes -- for the foreseeable future. 

Do I understand correctly that when a camera supports adjustable intervals, a MDA (with sequencable channels and/or piezo) drives the camera in streaming mode with the exposure defined by the channel’s exposure and the interval defined in the MDA delay?
What are the camera that support this feature? Would it be in principle possible to implement it with a Flash4 (by enhancing the device adapter and/or the driver) or is it beyond the camera specs?

> > Also, a new member of our group used to be able to acquire stacks faster (using another Flash4, maybe with more recent firmware…). He's using MM from Matlab (with snapImage() and getImage()) and is currently experiencing the same delays described in my previous email (while he used to be able to acquire 30ms images every 100ms, and move the stage between 2 images).
> > Is there any setting that we could have messed up in our  config file ?
> 
> I can't think of anything, other than difference in the camera version/firmware/settings. 

We’ve done further testing with this… When we run the following bsh script with 30ms exposure, the average time is ≈330ms.
>  for( int i =0; i<10; ++i){
>       start = System.currentTimeMillis();
>       mmc.snapImage();
>       mmc.getImage();
>       print(System.currentTimeMillis() - start);
>    }
This is very puzzling because when we click as fast as possible on the “Add to Album” button of the GUI, we reach almost 10Hz!! So somehow MM And the camera can talk faster to each other…
Also we were able to test that this doesn’t come from our old firmware (2.03A) since Andreas Durandi (from Hamamatsu Switzerland) was kind enough to configure another flash4 (firmware 2.50A) with MM1.4 and run the same script: he obtains similar delays (≈350ms).

So I don’t understand what’s happening and what prevents us to operate the camera faster in capture mode. Any explanation / hint would be very much appreciated!
Best,
Thomas

--
Thomas Julou  |  Computational & Systems Biology  |  Biozentrum – University of Basel  |  Klingelbergstrasse 50/70 CH-4056 Basel  |  +41 (0)61 267 16 21





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