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<span>Dear Jan,<br><br>Thanks for your comments. I have already available some ND filters, so I will set the laser<br>
power to the minimum and make some tests. I will determine my FOV and then to buy<br>
the Ruler Slides accordingly if necessary. I think I will try first the method that Jan Krieger<br>
and Alexis Maizel used with a scratched little mirror, (seems to be cheaper and very good too).<br>
<br>Kind regards,<br>Edgar<br><table border="0" cellpadding="0" cellspacing="0" width="457,5">
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<br>Fraunhofer-Institut für Produktionstechnologie IPT <br>Edgar Escobar Nieto <br>
<br><br> <br> <br>Steinbachstraße 17 <br>52074 Aachen <br><br>edgar.escobar.nieto@ipt.fraunhofer.de </font>
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<font color="blue" face="sans-serif" size="2"><u>http://www.ipt.fraunhofer.de</u>
</font></a><font face="sans-serif" size="2"><br>_________________________________________________________________________</font>
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<br><br><font color="#990099">-----Jan Huisken <huisken@mpi-cbg.de> schrieb: -----</font>
<div style="padding-left:5px;"><div style="padding-right:0px;padding-left:5px;border-left:solid black 2px;">
An: edgar.escobar.nieto@ipt.fraunhofer.de<br>Von: Jan Huisken <huisken@mpi-cbg.de><br>
Datum: 06.08.2013 23:21<br>Kopie: openspim@openspim.org<br>Betreff: Re: [OpenSPIM] About the calibration of the lightsheet<br>
<br>Hi Edgar<div><br><div><blockquote type="cite"><font face="Default Sans Serif,Verdana,Arial,Helvetica,sans-serif" size="2">
<span><table border="0" cellpadding="0" cellspacing="0" width="457,5"><tbody>
</tbody></table></span><font color="#990099">-----Weitergeleitet von Edgar Escobar Nieto/Fraunhofer IPT am 05.08.2013 11:54 -----</font>
<div style="padding-left:5px;"><div>An: <a href="mailto:openspim@openspim.org">
openspim@openspim.org</a><br>Von: Edgar Escobar Nieto/Fraunhofer IPT<br>
Datum: 31.07.2013 15:02<br>Betreff: (Unbenannt)<br><br><font face="Default Sans Serif,Verdana,Arial,Helvetica,sans-serif" size="2">
<span>Hi dear all,<br><br>I reached to the point where I need to calibrate the light-sheet. And I was wondering<br>
what is the optical density of the ND filter recommended to perform the calibration.<br>
</span></font></div></div></font></blockquote><div><br></div><div>Since you are reflecting the laser right onto the camera you need to be sure that you can turn down the laser intensity far enough that you do not damage your camera. Therefore the ND depends on the lowest power you can set your laser to and the sensitivity of your camera. The ND filter can be introduced in the illumination or detection path. Some fluorescence emission filter may also do the job.</div>
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<div style="padding-left:5px;"><div><font face="Default Sans Serif,Verdana,Arial,Helvetica,sans-serif" size="2">
<span><br>I would like to know also what is the </span><span>Frequency (lp/mm) recommended for the Opal Glass Ronchi Ruling Slides.<br>
I think that high a frequency would be better, but there must be a limit.<br>
</span></font></div></div></font></blockquote><div><br></div><div>The grid is used to (1) have some features on which you can focus and (2) to have some reflective parts the give you a profile of the beam. The distance does not matter very much. It is usually good if you have ca. 10 reflective lines across the entire FOV, which depends on your magnification, camera, etc. </div>
<div>In addition it is nice if you also have an adjacent region on this piece of glass that is fully reflective where you can inspect the full beam profile (after focusing on it with the grid part). Here you can align for sheet thickness and uniformity.</div>
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<span><br>Only to be sure, what I understood is that these Ruling Slides have already a reflective surface, so there is no<br>
need to glue a mirror to the ruling slides.<br></span></font></div></div>
</font></blockquote><div><br></div><div>No. The whole piece needs to be fairly thin to fit between the lenses. So you would have to cut out a small piece anyway. Cutting is not so easy but the cut does not need to be precise. Just smash it into pieces and pick the best piece ;-)</div>
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<div style="padding-left:5px;"><div><font face="Default Sans Serif,Verdana,Arial,Helvetica,sans-serif" size="2">
<span><br>Thank you in advance for your comments.<br></span></font></div>
</div></font></blockquote><div><br></div><div>Sure.</div><div><br></div>
<div>Best</div><div>Jan</div><div><br></div><div><div style="font-size: 11px; ">
Dr. Jan Huisken</div><div style="font-size: 11px; "><b>MPI of Molecular Cell Biology and Genetics</b>
</div><div style="font-size: 11px; ">Pfotenhauerstr. 108, 01307 Dresden, Germany</div>
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<span><br>Kind regards,<br>Edgar Escobar Nieto<br><table border="0" cellpadding="0" cellspacing="0" width="457,5">
<tbody><tr><td colspan="3"><font face="sans-serif" size="2">_________________________________________________________________________<br>
<br>Fraunhofer-Institut für Produktionstechnologie IPT <br>Edgar Escobar Nieto <br>
<br><br> <br> <br>Steinbachstraße 17 <br>52074 Aachen <br><br><a href="mailto:edgar.escobar.nieto@ipt.fraunhofer.de">
edgar.escobar.nieto@ipt.fraunhofer.de</a> </font><font color="blue" size="3">
<u><br></u></font><a href="http://www.ipt.fraunhofer.de/"><font color="blue" face="sans-serif" size="2">
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