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<div>Thank you Fredrik and Peter! Right now I am looking at either an Olympus U-AEC 1x/2x magnification changer or else the DE20BXC 2x magnifying camera adapter from SPOT Imaging. </div>
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<div>I checked the Nyquist sampling frequency for deconvolution at www.svi.nl/NyquistCalculator, and it looks like magnification is a good idea. The optimal sampling frequency for widefield with a 0.5 NA water lens is 0.26 um per px. I am imaging at 0.33
um/px with a 1x camera adapter. Therefore I have a little resolution to gain, though a 2x adapter at 0.16 um/px will be grossly oversampling. It might be a good idea to use a 1.5x adapter instead, as 0.22 um/px is pretty close to the theoretical limit. As
I understand it SPIM improves the axial resolution of images but does not change lateral resolution relative to widefield. Is this correct? </div>
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<div>All the best, </div>
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<div>Tim</div>
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<div>Timothy Feinstein, Ph.D. </div>
<div>Research Scientist</div>
<div>University of Pittsburgh Department of Developmental Biology</div>
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<span style="font-weight:bold">From: </span>Fredrik Ek <<a href="mailto:fredrik.ek@med.lu.se">fredrik.ek@med.lu.se</a>><br>
<span style="font-weight:bold">Date: </span>Wednesday, September 23, 2015 at 4:34 AM<br>
<span style="font-weight:bold">To: </span>Timothy Feinstein <<a href="mailto:tnf8@pitt.edu">tnf8@pitt.edu</a>>, openspim <<a href="mailto:openspim@openspim.org">openspim@openspim.org</a>><br>
<span style="font-weight:bold">Subject: </span>SV: changing "objectives"<br>
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<p class="MsoNormal"><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Hi,
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">We are using a Olympus magnifier 1x,2x and 4x with c-mount together with a Flash4.0 v2 and it works very well with the SPIM setup (20x 0.5 objective).
<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Best,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Fredrik<o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size: 10pt; font-family: Arial, sans-serif; color: black;"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size: 10pt; font-family: Arial, sans-serif; color: black;">Fredrik Ek, PhD,
</span><span style="font-size: 10pt; font-family: Tahoma, sans-serif; color: black;"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size: 10pt; font-family: Arial, sans-serif; color: black;">Researcher, Chemical Biology & Therapeutics
</span><span style="font-size: 10pt; font-family: Tahoma, sans-serif; color: black;"><br>
</span><span style="font-size: 10pt; font-family: Arial, sans-serif; color: black;">Department of Experimental Medical Science
</span><span style="font-size: 10pt; font-family: Tahoma, sans-serif; color: black;"><br>
</span><span style="font-size: 10pt; font-family: Arial, sans-serif; color: black;">Lund University, BMC D10
</span><span style="font-size: 10pt; font-family: Tahoma, sans-serif; color: black;"><br>
</span><span style="font-size: 10pt; font-family: Arial, sans-serif; color: black;">S-221 84 Lund, SWEDEN
</span><span style="font-size: 10pt; font-family: Tahoma, sans-serif; color: black;"><br>
</span><span style="font-size: 10pt; font-family: Arial, sans-serif; color: black;">------------------------
</span><span style="font-size: 10pt; font-family: Tahoma, sans-serif; color: black;"><br>
</span><span lang="SV" style="font-size:10.0pt;font-family:"Tahoma","sans-serif";color:black"><a href="mailto:Fredrik.Ek@med.lu.se"><span lang="EN-US" style="font-family: Arial, sans-serif; color: blue;">Fredrik.Ek@med.lu.se</span></a></span><span lang="SV" style="font-size: 10pt; font-family: Arial, sans-serif; color: black;"></span><span style="font-size: 10pt; font-family: Tahoma, sans-serif; color: black;"><br>
</span><span style="font-size: 10pt; font-family: Arial, sans-serif; color: black;">Phone: +46-727 17 22 05
</span><span style="font-size: 10pt; font-family: Tahoma, sans-serif; color: black;"><o:p></o:p></span></p>
<p class="MsoNormal"><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span lang="SV" style="font-size: 10pt; font-family: Tahoma, sans-serif;">Från:</span></b><span lang="SV" style="font-size: 10pt; font-family: Tahoma, sans-serif;">
<a href="mailto:openspim-bounces@openspim.org">openspim-bounces@openspim.org</a> [<a href="mailto:openspim-bounces@openspim.org">mailto:openspim-bounces@openspim.org</a>]
<b>För </b>Feinstein, Timothy N<br>
<b>Skickat:</b> den 22 september 2015 17:35<br>
<b>Till:</b> openspim<br>
<b>Ämne:</b> [OpenSPIM] changing "objectives"<o:p></o:p></span></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;">Hi folks, <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;">Thanks to the indispensable help of many users here, our SPIM is up and working. You are all the best. <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;">Since a SPIM makes it quite hard to change objectives, I bought 1x and a 0.5x C-mount camera adapters to get the equivalent of a 20x and a 10x objective. It
seems to work quite well. Now I wonder whether anyone has tried a magnifying tube lens like the Olympus U-CA or U-ECA to get a higher power image. It all depends on whether a 'vanilla' openSPIM design is already imaging near the Nyquist limit. We get 0.33
um/px using the Olympus 20x/0.5 and a Hamamatsu Flash4.0 v2, and even with deconvolution it may be pointless to magnify the image any further. Any thoughts appreciated. <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;">Alternatively, has anyone tried keeping a second sample chamber with a higher power objective to swap in as needed? It seems like a pain but less cumbersome
than building a second instrument. <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;">Thanks, <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;">Tim<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;">Timothy Feinstein, Ph.D. <o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;">Research Scientist<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;">University of Pittsburgh Department of Developmental Biology<o:p></o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;"><o:p> </o:p></span></p>
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<p class="MsoNormal"><span style="font-size: 10.5pt; font-family: Calibri, sans-serif; color: black;"><o:p> </o:p></span></p>
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