Combining z stack with timelapse

Posted by Julia Edgar on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Combining-z-stack-with-timelapse-tp7589566.html

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Thanks Timothy.
I have now changed the subject header.....
I'm imaging the movement of organelles (~ 50 nm diameter) within a structure with a depth of around 2-3 um, hence the reason I need to combine timelapse (1 frame every 10 seconds) with z stack.
Best wishes

Julia


As a rule I try to keep the analysis and the presentation as straightforward as possible by reducing dimensions and cropping. If I have to include depth, time, and color then I usually generate a volume projection and have it rotate slowly while advancing in time. Most proprietary scope software is good at this, though you often have to pay a substantial extra license for the volume/movie option. ImageJ/Fiji is not ideal but I  found Fluorender and Vaa3D to be good free alternatives.

Bear in mind that volume rendering involves a lot of semi-arbitrary and nonlinear transformations.  So that sort of display is best if it can supplement quantitative analysis rather than stand on its own.

Best,


T

Timothy Feinstein, Ph.D.


> On May 25, 2019, at 3:49 AM, Julia Edgar <[hidden email]> wrote:
>
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> *****
>
> Dear All
> A very naive question I know...
>
> How do you display time-lapse combined with confocal z stack? Flatten the z stack and play the time-lapse movie of the flattened (maximum intensity projection?) images?
>
> If so, how do you generate the movie using Zen software and/or Fiji?
>
> Many thanks in advance.
> Julia
>
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> ________________________________
> From: Confocal Microscopy List <[hidden email]> on behalf of Ralf Palmisano <[hidden email]>
> Sent: Saturday, May 25, 2019 1:47 am
> To: [hidden email]
> Subject: Re: SSD-RAID for new wide-field microscopy system
>
> *****
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> Hi Mirco,
>
> I am literally signing everything Nuno Mureno wrote. In addition whoever is the manufatcurer. I consider it bad advise to invest in an 11 GB graphics card and even worse to call 6.000$ for a 2 TB SSD extension. That is just playing foul or they have no clue about what they are selling and why...
>
> My pennie in here
>
> Ralph
>
>> Am 23/05/2019 um 14:57 schrieb Nuno Moreno:
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>> *****
>>
>> Hi Mirco
>>
>> Honestly I see no reason to have so much memory and such a high spec graphics card for an acquisition computer. Regarding the HDs, I would keep the 2TB and add extra space non SSD. Then just automate data flow from the fast to the slow drive. 2TB extra for 6000$ is just too much and might be too short anyway.
>>
>> Just my 2 cents
>> \N
>>
>>
>>
>>> On 23 May 2019, at 14:38, Mirco Martino <[hidden email]> wrote:
>>>
>>> *****
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>>>
>>> Hi all,
>>>
>>> we are buying a new fluorescent microscope wide-field system. The system include a workstation with 64GB DDR4 RAM, 11GB of graphic card, 512 gb of memory and SSD-RAID of 2TB. There is an option to increase the SSD-RAID to 4TB for about 6000$ extra. The system will be use for both fixed materiel imaging and for live-imaging with time-laps, z-stack, tiling and multi-fluorophores experiments. I was wondering if it is worth to invest those extra money for this option or not. This will be the first time we will have time-laps experiments, so I don't know what to expect as pictures size.
>>>
>>> What do you think?
>>>
>>> Thanks in advance for your comments and opinions!
>>>
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