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<div><span style="font-family: Calibri; font-size: 11pt; font-weight: bold;">Date:
</span><span style="font-family: Calibri; font-size: 11pt;">Friday, October 2, 2015 at 2:16 PM</span></div>
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<span style="font-weight:bold">To: </span>Michael Weber <<a href="mailto:weber@mpi-cbg.de">weber@mpi-cbg.de</a>><br>
<span style="font-weight:bold">Subject: </span>Re: [OpenSPIM] Alignment / Calibration Slides<br>
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<div>Regarding cutting that glass slide, I found the diamond wheel attachment on a Dremel tool to be indispensable for that. I was able to grind the slide fragment to a skinny trapezoid and wedge it into the opening of a 1 ml syringe without using glue. </div>
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<div>T <br>
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Timothy Feinstein, Ph.D.
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On Oct 2, 2015, at 10:22 AM, Michael Weber <<a href="mailto:weber@mpi-cbg.de">weber@mpi-cbg.de</a>> wrote:<br>
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<div><base href="x-msg://15/">Dear Neil,
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<div>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:</div>
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<div>Edmund Optics "Multi-Grid Standard Stage Micrometer"</div>
<div><a href="http://www.edmundoptics.com/test-targets/image-analysis-test-targets/multi-grid-standard-stage-micrometer/2700/">http://www.edmundoptics.com/test-targets/image-analysis-test-targets/multi-grid-standard-stage-micrometer/2700/</a></div>
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<div>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.</div>
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<div>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.</div>
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<div>Best,</div>
<div>Michael</div>
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<div>On Oct 1, 2015, at 6:35 PM, "Anthony, Neil" <<a href="mailto:nantho2@EMORY.EDU">nantho2@EMORY.EDU</a>> wrote:</div>
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Hi all,<o:p></o:p></div>
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<o:p> </o:p></div>
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I was just reading the paper suggested by Jan in Aug (<span lang="EN-GB">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.<o:p></o:p></span></div>
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<span lang="EN-GB"> </span></div>
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<span lang="EN-GB">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.<o:p></o:p></span></div>
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<span lang="EN-GB">Thanks<o:p></o:p></span></div>
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<span lang="EN-GB">Neil<o:p></o:p></span></div>
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<o:p> </o:p></div>
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<b><span style="font-size: 11.5pt; font-family: Tahoma, sans-serif;">Neil Anthony, PhD</span></b><span style="font-size: 11.5pt; font-family: Tahoma, sans-serif;"> | Research Imaging Specialist<br>
</span><span style="font-size: 10pt; font-family: Tahoma, sans-serif;">Health Sciences Research Building (HSRB), Room EG21<br>
1760 Haygood Drive, Atlanta, GA 30322</span><span style="font-size: 12pt; font-family: 'Times New Roman', serif;"><o:p></o:p></span></div>
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<span style="font-size: 10.5pt; font-family: Tahoma, sans-serif;"><a href="mailto:neil.anthony@emory.edu" style="color: purple; text-decoration: underline; "><span style="color: blue; ">neil.anthony@emory.edu</span></a><br>
<a href="http://www.cores.emory.edu/ici/" style="color: purple; text-decoration: underline; "><span style="color: blue; ">ici.emory.edu</span></a></span><span style="font-size: 12pt; font-family: 'Times New Roman', serif;"><br>
404 969-CORE<o:p></o:p></span></div>
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<a href="http://www.cores.emory.edu/ici/" style="color: purple; text-decoration: underline; "><span style="font-size: 12pt; font-family: 'Times New Roman', serif; color: blue; text-decoration: none;"><image001.png></span></a><span style="font-size: 12pt; font-family: 'Times New Roman', serif;"><o:p></o:p></span></div>
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<a href="http://www.linkedin.com/pub/neil-anthony/35/a0b/126" style="color: purple; text-decoration: underline; "><span style="font-size: 12pt; font-family: 'Times New Roman', serif; color: blue; text-decoration: none;"><image003.png></span></a><span style="font-size: 12pt; font-family: 'Times New Roman', serif;"><o:p></o:p></span></div>
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<a href="https://www.researchgate.net/profile/Neil_Anthony/" style="color: purple; text-decoration: underline; "><span style="font-size: 12pt; font-family: 'Times New Roman', serif; color: blue; text-decoration: none;"><image004.png></span></a><span style="font-size: 12pt; font-family: 'Times New Roman', serif;"><o:p></o:p></span></div>
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<o:p> </o:p></div>
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_____________<br>
<br>
Michael Weber<br>
Postdoc, Huisken lab<br>
Max Planck Institute of Molecular Cell Biology and Genetics<br>
Pfotenhauerstrasse 108, 01307 Dresden<br>
Tel. 0049 351/2102837<br>
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<a href="http://www.mpi-cbg.de/huisken">http://www.mpi-cbg.de/huisken</a></div>
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