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<span>Hi Jan,<br><br>I changed the order of the lens in the telescope so that it expands the beam and I got a thinner lightsheet, but when I move the mirror mounted on the sample holder in the X direction<br>
the lightsheet expands too much. I attach some images.<br><br>Do you know if I can adjust the relationship between the thickness of the lightsheet in the center and in the borders of the FOV using the vertical slit?<br>
<br>Kind regards,<br>Edgar<br><br><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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<br><br><font color="#990099">-----Weitergeleitet von Edgar Escobar Nieto/Fraunhofer IPT am 05.12.2013 15:53 -----</font>
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An: Jan Krieger <j.krieger@dkfz-heidelberg.de><br>Von: Edgar Escobar Nieto/Fraunhofer IPT<br>
Datum: 04.12.2013 17:37<br>Betreff: Re: [OpenSPIM] About the lightsheet<br>
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<span>Hi Jan,<br><br>Yes, I am talking about the telescope between the corner mirror and the objective.<br>
<br>Ohh, I have thought that this telescope instead of expanding the beam, it was reducing it.<br>
In the instructions of how to install the telescope from the OpenSPIM wiki, the 50 mm focal length lens goes first and then<br>
the 25 mm, (the opposite order in comparisson to the beam expander). So, it looks like all this time I have been using the<br>
wrong lens configuration. <br><br>Concerning the pictures, I am always using the illumination objective, in some using the telescope (between the corner mirror and the objective), in others without using it.<br>
<br>That explain everything. I will change the order of the lenses of the last telescope in order to expand the beam and not to reduce it. Thanks a lot for your answer.<br>
I will let you know if I suceed.<br><br>Kind regards,<br>Edgar<br><br><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>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 Krieger <j.krieger@dkfz-heidelberg.de> schrieb: -----</font>
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An: "edgar.escobar.nieto@ipt.fraunhofer.de" <edgar.escobar.nieto@ipt.fraunhofer.de><br>
Von: Jan Krieger <j.krieger@dkfz-heidelberg.de><br>Datum: 04.12.2013 15:39<br>
Betreff: Re: [OpenSPIM] About the lightsheet<br><br><div><font face="Courier New,Courier,monospace" size="2">
Hi!<br><br>hhmmm ... I pressume you're talking about the telescope between the <br>
corner mirror and the objective, right?<br><br>As far as I know the openSPIM, this telescope should lead to an <br>
additional 2-fold expansion of the beam. This means that the light fills <br>
more of the projection objective and should thus also create a thinner <br>
focus (we "use more" of the available NA of the objective ... ) ... So <br>
your finding of with and without the objective looks strange to me ... <br>
Can you measure the width of the lightsheet with your different <br>configurations? It should slightly expand, if you move the 45°-mirror <br>
along the lightsheet axis ... How did you find the right position of the <br>
projection objective? We usually find a sharp line on the mirror and <br>
position it in the center of the FOV, then adjust the lightsheet to <br>
overlay with this line, using the corner mirror and finally adjust the <br>
projection obvjective (along the lightsheet axis) until we have the <br>
thinnest possible lightsheet which should then also have it's thinnest <br>
position in the center of the FOV.<br><br>Best,<br>JAN<br><br><br><br>Am 04.12.2013 15:18, schrieb edgar.escobar.nieto@ipt.fraunhofer.de:<br>
> Hi dear all,<br>><br>> I have made some tests about the lightsheet. I made some pictures of the<br>
> lightsheet using the telescope and without using it. To be able of<br>
> imaging the lightsheet I used a small mirror at 45° mounted in a sample<br>
> holder to direct the lightsheet onto the CCD.<br>><br>> I found that the lightsheet looks narrower without using the telescope.<br>
> I know that the telescope is used for alignig the lightsheet with the<br>
> focal plane of the detection objective in<br>> an easier way (adjusting the knob of the last mirror causes the<br>
> lightsheet to move parallel to the plane of the detection objective<br>
> without being tilted).<br>><br>> I found out that also is possible to align the lightsheet without using<br>
> the telescope. Then, I aligned the lightsheet rotating the knob of the<br>
> mirror but also translating it (I have mounted the mirror in a rail as<br>
> in this picture <a href="http://openspim.org/File:1I_1D_OpenSPIM_03.jpg">
http://openspim.org/File:1I_1D_OpenSPIM_03.jpg</a> ). To be<br>> sure that the lightsheet was perpendicular to the detection axis<br>
> throughout the complete FOV I made some pictures of the lightsheet<br>
> moving the mirror mounted in the sample holder in the X direction.<br>
><br>> I attach the pictures to this mail. In the pictures can be observed that<br>
> the lightsheet is directed to the part of the mirror mounted in the<br>
> sample holder that is in focus. Even if I translate the mirror mounted<br>
> in the sample holder in the X direction, the lightsheet remains on the<br>
> focused part. This is the way I check that the lightsheet is<br>> perpendicular throughout the complete FOV. When the lightsheet was not<br>
> completely perpendicular I observed that the lightsheet was not<br>
> reflected exactly on the part in focus when moving the mirror in the X<br>
> direction.<br>><br>> From the pictures without using the telescope, it can be observed that<br>
> the lightsheet widens in the borders of the FOV (because of the gaussian<br>
> nature of the lightsheet), but in the pictures where I use the<br>
> telescope, the lightsheet seems to be the same thickness along the<br>
> complete FOV.<br>><br>> I would like to know if what I have done is right and why does the<br>
> lightsheet does not seem to change its thickness in the borders of the<br>
> FOV when I use the telescope.<br>><br>> Kind regards,<br>> Edgar<br>
> _________________________________________________________________________<br>
><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 _<br>> __http://www.ipt.fraunhofer.de_ <<a href="http://www.ipt.fraunhofer.de/">
http://www.ipt.fraunhofer.de/</a>><br>> _________________________________________________________________________<br>
><br>> <br>> <br>><br><br></font></div></div>
<br><br>[Anhang 'j_krieger.vcf' entfernt von Edgar Escobar Nieto/Fraunhofer IPT]</div>
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