Brainbow on Zeiss Lightsheet Z1 anyone?

[Holly L. AARON]

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Hi, there -

Is anyone out there imaging Brainbow specimens (we are planning to have
cleared mice brains soon....) with the Zeiss Lightsheet Z1? Our filter
choices do not seem to be ideal for it and we are looking to see what
filters would be recommended and if we can get them in place ASAP. Filter
info below.

Thanks so much!

Available Lasers: 405, 445, 488, 514, 561, and 640nm
Laser blocking filters (these cannot be changed during an experiment,
ideally):

   - 405/488/561/640
   - 445/514/561

Emission Filters (again, we can use either FW1 or FW2, but cannot really
change during an experiment... at least, not easily... ):

> FW1:
> - LP510, BP 420 - 470, BP 575 - 615
> - LP 510, BP 460-500, BP 525 - 565
> - LP560, BP 505 - 545, BP 575-615
> - LP560, BP 505 - 545, LP 585
> - LP 560, BP 505 - 545, LP 660
> FW2:
> - BP 460 - 500, LP 525 - 565
> - BP 460 - 500, LP 585
> - BP 525 - 565, LP 585

Cheers,
 Holly

*Holly Aaron, [hidden email] <[hidden email]>*
*CRL Molecular Imaging Center*

Register now for AIM:
*Advanced Imaging Methods Workshop*
*January 24 – 26, 2018*
*https://www.regonline.com/AIM2018* <https://www.regonline.com/AIM2018>

[Douglas Richardson]

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Hi Holly,



Be careful with brainbow.  There are so many versions of it out there
now.  Make
sure you know exactly which FPs are in the system.  Sometimes I make people
prove it to me by taking a spectral image first (on a confocal or
widefield).



If it’s Brainbow 1.0 (CFP, YFP, RFP) you’ll likely be able to get away with
the stock green/red cube with 488 and 561 lasers and the stock CFP cube
with the 445 laser.  You can get away with a little bleed through between
channels, but too much makes it impossible to get anything other than
pastels or white when you merge the channels.



Other brainbow versions (or if you have bleed through) may require some
custom filters from chroma/semrock.  These are easy to swap yourself on the
lightsheet system, you just have to remember to realign the cameras for all
cubes when you put the wheel back into the system.



-Doug

On Fri, Oct 6, 2017 at 2:37 PM, Holly L. AARON <[hidden email]> 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.
> *****
>
> Hi, there -
>
> Is anyone out there imaging Brainbow specimens (we are planning to have
> cleared mice brains soon....) with the Zeiss Lightsheet Z1? Our filter
> choices do not seem to be ideal for it and we are looking to see what
> filters would be recommended and if we can get them in place ASAP. Filter
> info below.
>
> Thanks so much!
>
> Available Lasers: 405, 445, 488, 514, 561, and 640nm
> Laser blocking filters (these cannot be changed during an experiment,
> ideally):
>
>    - 405/488/561/640
>    - 445/514/561
>
> Emission Filters (again, we can use either FW1 or FW2, but cannot really
> change during an experiment... at least, not easily... ):
>
> > FW1:
> > - LP510, BP 420 - 470, BP 575 - 615
> > - LP 510, BP 460-500, BP 525 - 565
> > - LP560, BP 505 - 545, BP 575-615
> > - LP560, BP 505 - 545, LP 585
> > - LP 560, BP 505 - 545, LP 660
> > FW2:
> > - BP 460 - 500, LP 525 - 565
> > - BP 460 - 500, LP 585
> > - BP 525 - 565, LP 585
>
>
> Cheers,
>  Holly
>
> *Holly Aaron, [hidden email] <[hidden email]>*
> *CRL Molecular Imaging Center*
>
> Register now for AIM:
> *Advanced Imaging Methods Workshop*
> *January 24 – 26, 2018*
> *https://www.regonline.com/AIM2018* <https://www.regonline.com/AIM2018>
>