[OpenSPIM] Alignment / Calibration Slides

[Michael Weber]

Dear Neil,

hope you enjoyed reading the paper :). We use a home-made mirror/grid for aligning our setups. The "mirror/grid" part is cut from a stage micrometer:

Edmund Optics "Multi-Grid Standard Stage Micrometer"
http://www.edmundoptics.com/test-targets/image-analysis-test-targets/multi-grid-standard-stage-micrometer/2700/

Cutting the glass is tricky - maybe you have access to a workshop. You want to have both mirror and grid on top of each other on the same piece of glass, such that you just have to move the sample up and down to switch between them. One end of this piece of glass is then mounted in the tip of a 1 ml Braun Omnifix-F syringe using glue. That's the second difficult step. Ideally, the glass surface ends up straight in the rotation axis of your setup. Again, if you have access to a workshop, they might be able to help you with that. The syringe is then mounted in the sample holder. Make sure that the syringe has the right length for positioning the glass in the focal plane. One can also think of a machined or 3D-printed adapter (or entire holder) for the piece of glass.

Another tool that is really useful for alignment are fluorescent beads / microspheres in 1.5% agarose. This is especially good for the fine tuning of the light sheet position, PSF measurements and so on. Mirror/grid are more suitable for a coarse alignment.

Best,
Michael


On Oct 1, 2015, at 6:35 PM, "Anthony, Neil" <nantho2 at EMORY.EDU> wrote:

> Hi all,
>  
> I was just reading the paper suggested by Jan in Aug (Weber, Michael, Michaela Mickoleit, and Jan Huisken. 2014. “Light Sheet Microscopy.” Methods in Cell Biology 123: 193–215. doi:10.1016/B978-0-12-420138-5.00011-2.), thinking ahead to the calibration/alignment part of the setup.
>  
> Does anybody have part numbers and/or websites for the required pieces?  Also, I’d be interested in how you’ve positioned and held them at the objective focus, and ensured their correct orientation.
>  
> Thanks
> Neil
>  
> Neil Anthony, PhD  | Research Imaging Specialist
> Health Sciences Research Building (HSRB), Room EG21
> 1760 Haygood Drive, Atlanta, GA 30322
> neil.anthony at emory.edu
> ici.emory.edu
> 404 969-CORE
> <image001.png>
> <image002.png>
> <image003.png>
> <image004.png>
>  
> 
> 
> This e-mail message (including any attachments) is for the sole use of
> the intended recipient(s) and may contain confidential and privileged
> information. If the reader of this message is not the intended
> recipient, you are hereby notified that any dissemination, distribution
> or copying of this message (including any attachments) is strictly
> prohibited.
> 
> If you have received this message in error, please contact
> the sender by reply e-mail message and destroy all copies of the
> original message (including attachments).
> _______________________________________________
> OpenSPIM mailing list
> OpenSPIM at openspim.org
> http://openspim.org/mailman/listinfo/openspim

_____________

Michael Weber
Postdoc, Huisken lab
Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108, 01307 Dresden
Tel. 0049 351/2102837

http://www.mpi-cbg.de/huisken

-------------- next part --------------
An HTML attachment was scrubbed...
URL: <http://openspim.org/pipermail/openspim/attachments/20151002/9c027943/attachment-0002.html>