FRAP, multipoint time series for 2 hours using Zen on Zeiss 510 LSM

[Michelle Peckham]

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Dear Listers

We've just upgraded the software on our Zeiss LSM 510 to Zen, and I now have to work out how to set up a macro to:
Select about 10-15 cells, and mark their positions (X,Y,Z)
Zoom in, set image capture parameters, and rotate cells of interest so they all have the same orientation
Place a rectangle (the bleach area) precisely on an specific region of each cell for FRAP
Take an image for each cell
FRAP the cells one by one
Collect images after frap for the next ~2 hours (t/12 is about 30 minutes for fluorescence recovery for my protein of interest) every 5 minutes

(I did have a macro that used to do most of this on the old LSM software, although it was an absolute pain to set up!)

I will be talking to the Zeiss reps tomorrow, but just wondered if anyone had any advice?

Cheers

Michelle

[DamirSudar]

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Hi Michelle,

Writing macros in both AIM and ZEN is a fairly complicated affair and
you'd need the license for the Zeiss VBA macro editor which will set you
back quite a few dollars. It may well be that Zeiss's MultiTime macro
(also not cheap) will actually do what you need and that will do a lot
of even more complex things if you get more ambitious later. So when
talking with Zeiss, check with them whether MultiTime will do the job.
It was developed by Zbi Iwinski from Zeiss USA and Zbi is a great
resource for all complex things you might want to do with macros and such.

Good luck,
- Damir

On 8/7/2012 11:24 AM, Michelle Peckham wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear Listers
>
> We've just upgraded the software on our Zeiss LSM 510 to Zen, and I now have to work out how to set up a macro to:
> Select about 10-15 cells, and mark their positions (X,Y,Z)
> Zoom in, set image capture parameters, and rotate cells of interest so they all have the same orientation
> Place a rectangle (the bleach area) precisely on an specific region of each cell for FRAP
> Take an image for each cell
> FRAP the cells one by one
> Collect images after frap for the next ~2 hours (t/12 is about 30 minutes for fluorescence recovery for my protein of interest) every 5 minutes
>
> (I did have a macro that used to do most of this on the old LSM software, although it was an absolute pain to set up!)
>
> I will be talking to the Zeiss reps tomorrow, but just wondered if anyone had any advice?
>
> Cheers
>
> Michelle

--
Damir Sudar - Staff Scientist and Deputy for Technology
Lawrence Berkeley Laboratory / Life Sciences Division
One Cyclotron Road, MS 977, Berkeley, CA 94720, USA
T: 510/486-5346 - F: 510/486-5586 - E: [hidden email]
WWW: http://www.lbl.gov/lsd/People_&_Organization/Scientific_Staff_Directory/Sudar_Lab.html

[Terclavers, Sven]

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Dear Michelle,

To run a multi-position experiment combined with the bleaching for FRAP,
different frame-orientation/zoom settings, time-lapse etc. , you can use
the MTS macro (MultiTime), which can be found in C:\ZEN\Macros.  Most
probably this was the macro you were using before, however it'll be
another version updated for ZEN.

With best regards,

Sven



From:
Michelle Peckham <[hidden email]>
To:
[hidden email]
Date:
07.08.12 20:27
Subject:
FRAP, multipoint time series for 2 hours using Zen on Zeiss 510 LSM
Sent by:
Confocal Microscopy List <[hidden email]>



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Dear Listers

We've just upgraded the software on our Zeiss LSM 510 to Zen, and I now
have to work out how to set up a macro to:
Select about 10-15 cells, and mark their positions (X,Y,Z)
Zoom in, set image capture parameters, and rotate cells of interest so
they all have the same orientation
Place a rectangle (the bleach area) precisely on an specific region of
each cell for FRAP
Take an image for each cell
FRAP the cells one by one
Collect images after frap for the next ~2 hours (t/12 is about 30 minutes
for fluorescence recovery for my protein of interest) every 5 minutes

(I did have a macro that used to do most of this on the old LSM software,
although it was an absolute pain to set up!)

I will be talking to the Zeiss reps tomorrow, but just wondered if anyone
had any advice?

Cheers

Michelle




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