<html><head><meta http-equiv="Content-Type" content="text/html charset=windows-1252"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space;" class="">Hey Neil,<div class=""><br class=""></div><div class="">valid questions…</div><div class=""><br class=""></div><div class=""><blockquote type="cite" class=""><div class="" style="background-color: rgb(255, 255, 255); font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; color: rgb(31, 73, 125);"><div class="">how the alignment of the cylindrical lens might affect things when you place it back in the path?</div></div></blockquote><div class=""><div class=""><br class="webkit-block-placeholder"></div><div apple-content-edited="true" class="">
<div style="color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-stroke-width: 0px;" class=""><div class="">Not much really… It doesn’t even seem to matter if the lens is closer or further away from the mirror by a full centimeter. BUT, I would still recommend putting it in focus/right place.<br class="Apple-interchange-newline"><br class=""></div><div class=""><blockquote type="cite" class=""><div class="" style="background-color: rgb(255, 255, 255); font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; color: rgb(31, 73, 125);"><div class="">how sensitive are things to it being off axis, not quite perpendicular, or slightly rotated?</div></div></blockquote></div><div class=""><br class=""></div><div class="">Not very much. But there are ways to find out if it is though, although this requires a bead sample and the emission filter needs to be replaced into the infinity tube. </div><div class=""> - For determining the rotation of the cylindrical lens, watch the top and bottom third of the field of view of the camera chip to make sure it is evenly illuminated.</div><div class=""> - For determining the “perpendicularity” of the sheet, watch the left and right third of the field of view of the camera chip to make sure it is evenly illuminated. Here though you have to adjust mirrors… not fun.</div><div class=""><br class=""></div><div class=""><blockquote type="cite" class=""><div class="" style="background-color: rgb(255, 255, 255); font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; color: rgb(31, 73, 125);"><div class="">Also, can you view these things via the camera using the scattered light too?</div></div></blockquote></div><div class=""><br class=""></div><div class="">Try, you’ll see how dirty your media is…</div><div class=""><br class=""></div><div class="">Best Regards,</div><div class="">Pete</div><div class=""><br class=""></div><div class="">Peter Gabriel Pitrone - DipRMS TechRMS FRMS</div><div class="">Light Sheet Fluorescence Microscopist and Imaging Specialist</div><div class="">for Dr. Pavel Tomancak's research group at the</div><div class="">Max Planck Institute of Molecular Cell Biology and Genetics</div><div class="">Pfotenhauerstraße 108</div><div class="">01307 Dresden, Saxony</div><div class="">Germany</div><div class=""><br class=""></div><div class=""><br class=""></div><div class=""><a href="http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html" class="">http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html</a> &</div><div class=""><a href="http://www.openspim.org" class="">http://www.openspim.org</a></div><div class=""><br class=""></div><div class="">"I KEEP six honest serving-men (they taught me all I knew); their names are What and Why and When and How and Where and Who." Rudyard Kipling - The Elephant's Child</div></div><div style="color: rgb(0, 0, 0); font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-stroke-width: 0px;" class=""><br class=""></div><br class="Apple-interchange-newline">
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<br class=""><div><blockquote type="cite" class=""><div class="">On Nov 17, 2015, at 1:28 PM, Anthony, Neil <<a href="mailto:nantho2@EMORY.EDU" class="">nantho2@EMORY.EDU</a>> wrote:</div><br class="Apple-interchange-newline"><div class=""><div style="font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255); font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; color: rgb(31, 73, 125);" class=""><div class="">Hi Pete, thanks for the info on the alignment.</div><div class=""><br class=""></div><div class="">Can I ask how the alignment of the cylindrical lens might affect things when you place it back in the path? You slide it to create the best focus on the conjugate mirror(?), but how sensitive are things to it being off axis, not quite perpendicular, or slightly rotated? Also, can you view these things via the camera using the scattered light too? </div><div class=""><br class=""></div><div class="">I apologise if these are silly questions; I've not aligned the system yet, just started bolting it down.</div><div class=""><br class=""></div><div class="">Thanks for your time</div><div class="">Neil</div><div class=""><br class=""></div><div id="signature-x" style="font-family: Calibri, Arial, Helvetica, sans-serif; font-size: 12pt; color: rgb(31, 73, 125);" class=""><div class=""></div></div></div><div id="quoted_header" style="font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: 14px; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255); clear: both;" class=""><br class=""><div style="border-style: solid none none; border-top-color: rgb(225, 225, 225); border-top-width: 1pt; padding: 3pt 0cm 0cm;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""><b class="">From:</b><span class="Apple-converted-space"> </span>Peter Gabriel Pitrone <<a href="mailto:pitrone@mpi-cbg.de" class="">pitrone@mpi-cbg.de</a>><br class=""><b class="">Sent:</b><span class="Apple-converted-space"> </span>Nov 11, 2015 10:45 AM<br class=""><b class="">To:</b><span class="Apple-converted-space"> </span>Windell, Dylan<br class=""><b class="">Cc:</b><span class="Apple-converted-space"> </span><a href="mailto:openspim@openspim.org" class="">openspim@openspim.org</a><br class=""><b class="">Subject:</b><span class="Apple-converted-space"> </span>Re: [OpenSPIM] Light Sheet Alignment Procedure (Was: 3D Reconstruction)<br class=""></span></div></div><br type="attribution" style="font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: 14px; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255);" class=""><div style="font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: 14px; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px; -webkit-text-stroke-width: 0px; background-color: rgb(255, 255, 255);" class="">Howdy Dylan,<div class=""><br class=""></div><div class="">Go ahead and use an ND filter, it shouldn’t effect the process much at all… Sometimes I use them as well when the signal is too intense, but one can’t see any beads when it is in place.<br class=""><div class=""><div class="" style="font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px;"><div class=""><br class="Apple-interchange-newline"><br class=""></div><div class="">Best Regards,</div><div class="">Pete</div><div class=""><br class=""></div><div class="">Peter Gabriel Pitrone - DipRMS TechRMS FRMS</div><div class="">Light Sheet Fluorescence Microscopist and Imaging Specialist</div><div class="">for Dr. Pavel Tomancak's research group at the</div><div class="">Max Planck Institute of Molecular Cell Biology and Genetics</div><div class="">Pfotenhauerstraße 108</div><div class="">01307 Dresden, Saxony</div><div class="">Germany</div><div class=""><br class=""></div><div class=""><br class=""></div><div class=""><a href="http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html" class="">http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html</a><span class="Apple-converted-space"> </span>&</div><div class=""><a href="http://www.openspim.org/" class="">http://www.openspim.org</a></div><div class=""><br class=""></div><div class="">"I KEEP six honest serving-men (they taught me all I knew); their names are What and Why and When and How and Where and Who." Rudyard Kipling - The Elephant's Child</div></div><div class="" style="font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px;"><br class=""></div><br class="Apple-interchange-newline"></div><br class=""><div class=""><blockquote type="cite" class=""><div class="">On Nov 11, 2015, at 4:32 PM, Windell, Dylan <<a href="mailto:dw388@exeter.ac.uk" class="">dw388@exeter.ac.uk</a>> wrote:</div><br class="Apple-interchange-newline"><div class=""><div class="WordSection1" style="font-family: Helvetica; font-size: 12px; font-style: normal; font-variant: normal; font-weight: normal; letter-spacing: normal; line-height: normal; orphans: auto; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; widows: auto; word-spacing: 0px;"><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Hey Pete,</span></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Thanks for the instructions, I will try them out ASAP. I am guessing you meant replacing the cylindrical lens after the alignment?</span></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">My laser cannot be attenuated that low but if I were to use an ND filter, would that affect the alignment process?</span></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Kind regards,</span></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Dylan</span></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div><div class=""><div class="" style="border-style: solid none none; border-top-color: rgb(181, 196, 223); border-top-width: 1pt; padding: 3pt 0cm 0cm;"><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><b class=""><span lang="EN-US" class="" style="font-size: 10pt; font-family: Tahoma, sans-serif;">From:</span></b><span lang="EN-US" class="" style="font-size: 10pt; font-family: Tahoma, sans-serif;"><span class="Apple-converted-space"> </span>Peter Gabriel Pitrone [<a href="mailto:pitrone@mpi-cbg.de" class="">mailto:pitrone@mpi-cbg.de</a>]<span class="Apple-converted-space"> </span><br class=""><b class="">Sent:</b><span class="Apple-converted-space"> </span>11 November 2015 15:22<br class=""><b class="">To:</b><span class="Apple-converted-space"> </span>Windell, Dylan<br class=""><b class="">Cc:</b><span class="Apple-converted-space"> </span><a href="mailto:openspim@openspim.org" class="">openspim@openspim.org</a><br class=""><b class="">Subject:</b><span class="Apple-converted-space"> </span>Re: [OpenSPIM] Light Sheet Alignment Procedure (Was: 3D Reconstruction)</span></div></div></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div><p class="MsoNormal" style="margin: 0cm 0cm 12pt; font-size: 12pt; font-family: 'Times New Roman', serif;">Of course you need to replace the Cylindrical lens to SPIMage again… Sent that last one too fast.</p><div class=""><blockquote class="" style="margin-top: 5pt; margin-bottom: 5pt;"><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">On Nov 11, 2015, at 4:14 PM, Peter Gabriel Pitrone <<a href="mailto:pitrone@mpi-cbg.de" class="" style="color: purple; text-decoration: underline;">pitrone@mpi-cbg.de</a>> wrote:</div></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">Hey Dylan,</div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">I’ve started using a different technique for alignment that doesn’t require any thing at all (except for the media) in the sample chamber. I’ll describe it in short below:</div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">- Either take out the sample, or move it well beyond the focal plane (far away from the detection objective)</div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">- Take out all the filters blocking the laser from the camera (make sure that the laser is modulated way down to around ±1 mW)</div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">- Take out the cylindrical lens(es)</div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">- Go “Live” at a short exposure time (10-25 ms)</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">You should see an in-focus beam in the middle of the field of view that cuts across the image, and it should slightly converge and then diverge with the narrowest point in the middle of the field of view. Now you can adjust the conjugate plane (corner) mirror horizontally to correct any misalignment in terms of focus, and vertically to place it in the center across the field. The two lens telescope between the mirror and the illumination lens can be adjusted along the rail so the narrowest part of the beam is centered in the exact middle.</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Best Regards,</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Pete</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Peter Gabriel Pitrone - DipRMS TechRMS FRMS</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Light Sheet Fluorescence Microscopist and Imaging Specialist</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">for Dr. Pavel Tomancak's research group at the</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Max Planck Institute of Molecular Cell Biology and Genetics</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Pfotenhauerstraße 108</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">01307 Dresden, Saxony</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Germany</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"><a href="http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html" class="" style="color: purple; text-decoration: underline;">http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html</a><span class="Apple-converted-space"> </span>&</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"><a href="http://www.openspim.org/" class="" style="color: purple; text-decoration: underline;">http://www.openspim.org</a></span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">"I KEEP six honest serving-men (they taught me all I knew); their names are What and Why and When and How and Where and Who." Rudyard Kipling - The Elephant's Child</span></div></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div><div class=""><blockquote class="" style="margin-top: 5pt; margin-bottom: 5pt;"><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">On Nov 10, 2015, at 5:57 PM, Windell, Dylan <<a href="mailto:dw388@exeter.ac.uk" class="" style="color: purple; text-decoration: underline;">dw388@exeter.ac.uk</a>> wrote:</div></div><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Hi Pete,</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">I have a cut micrometre slide stuck in a syringe which I use to check the laser sheet straightness as per OpenSPIM instructions followed by checking the embryos for image sharpness. I probably should be doing it with beads but I am more comfortable with an embryo as I am not 100% what an optimal bead scan would look like.</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Kind regards,</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Dylan</span></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div></div><div class=""><div class="" style="border-style: solid none none; border-top-color: rgb(225, 225, 225); border-top-width: 1pt; padding: 3pt 0cm 0cm;"><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><b class=""><span lang="EN-US" class="" style="font-size: 11pt; font-family: Calibri, sans-serif;">From:</span></b><span class="apple-converted-space"><span lang="EN-US" class="" style="font-size: 11pt; font-family: Calibri, sans-serif;"> </span></span><span lang="EN-US" class="" style="font-size: 11pt; font-family: Calibri, sans-serif;">Peter Gabriel Pitrone [<a href="mailto:pitrone@mpi-cbg.de" class="" style="color: purple; text-decoration: underline;">mailto:pitrone@mpi-cbg.de</a>]<span class="apple-converted-space"> </span><br class=""><b class="">Sent:</b><span class="apple-converted-space"> </span>10 November 2015 17:41<br class=""><b class="">To:</b><span class="apple-converted-space"> </span>Windell, Dylan <<a href="mailto:dw388@exeter.ac.uk" class="" style="color: purple; text-decoration: underline;">dw388@exeter.ac.uk</a>><br class=""><b class="">Cc:</b><span class="apple-converted-space"> </span>Pavel Tomancak <<a href="mailto:tomancak@mpi-cbg.de" class="" style="color: purple; text-decoration: underline;">tomancak@mpi-cbg.de</a>>;<span class="Apple-converted-space"> </span><a href="mailto:openspim@openspim.org" class="" style="color: purple; text-decoration: underline;">openspim@openspim.org</a><br class=""><b class="">Subject:</b><span class="apple-converted-space"> </span>Re: [OpenSPIM] 3D Reconstruction</span></div></div></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">Hello Dylan,</div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">Can you tell me your light sheet alignment procedure that you do, hopefully before each experiment, maybe it could help exclude that possibility from the equation… </div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div><div class=""><div class=""><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Best Regards,</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Pete</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Peter Gabriel Pitrone - DipRMS TechRMS FRMS</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Light Sheet Fluorescence Microscopist and Imaging Specialist</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">for Dr. Pavel Tomancak's research group at the</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Max Planck Institute of Molecular Cell Biology and Genetics</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Pfotenhauerstraße 108</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">01307 Dresden, Saxony</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">Germany</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"><a href="http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html" class="" style="color: purple; text-decoration: underline;"><span class="" style="color: purple;">http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html</span></a><span class="apple-converted-space"> </span>&</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"><a href="http://www.openspim.org/" class="" style="color: purple; text-decoration: underline;"><span class="" style="color: purple;">http://www.openspim.org</span></a></span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;">"I KEEP six honest serving-men (they taught me all I knew); their names are What and Why and When and How and Where and Who." Rudyard Kipling - The Elephant's Child</span></div></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 9pt; font-family: Helvetica, sans-serif;"> </span></div></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div><div class=""><blockquote class="" style="margin-top: 5pt; margin-bottom: 5pt;"><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">On Nov 10, 2015, at 5:20 PM, Windell, Dylan <<a href="mailto:dw388@exeter.ac.uk" class="" style="color: purple; text-decoration: underline;"><span class="" style="color: purple;">dw388@exeter.ac.uk</span></a>> wrote:</div></div></div><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Dear Pavel,</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Thank you for your email. I just ran the registrations again on a couple of datasets I had. They were taken at 60 degrees apart at 10 or 20x magnification. The logs correspond with the raw data I have uploaded on my onedrive if you want to have a look at those too:</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"><a href="http://1drv.ms/1M426qo" class="" style="color: purple; text-decoration: underline;"><span class="" style="color: purple;">http://1drv.ms/1M426qo</span></a></span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">I have purchased some wider borosilicate capillaries that might help in both having more volume for beads and ensuring the tube is dead straight. Zhung, you might find that helpful? I will be running tests on them very soon so would let you know. I will also try Vaa3D, thanks for the suggestion.</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Let me know if you need any more information.</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Thanks again,</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"><br class="">Dylan</span></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"> </span></div></div></div><div class=""><div class="" style="border-style: solid none none; border-top-color: rgb(181, 196, 223); border-top-width: 1pt; padding: 3pt 0cm 0cm;"><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><b class=""><span lang="EN-US" class="" style="font-size: 10pt; font-family: Tahoma, sans-serif;">From:</span></b><span class="apple-converted-space"><span lang="EN-US" class="" style="font-size: 10pt; font-family: Tahoma, sans-serif;"> </span></span><span lang="EN-US" class="" style="font-size: 10pt; font-family: Tahoma, sans-serif;">Pavel Tomancak [<a href="mailto:tomancak@mpi-cbg.de" class="" style="color: purple; text-decoration: underline;"><span class="" style="color: purple;">mailto:tomancak@mpi-cbg.de</span></a>]<span class="apple-converted-space"> </span><br class=""><b class="">Sent:</b><span class="apple-converted-space"> </span>10 November 2015 13:33<br class=""><b class="">To:</b><span class="apple-converted-space"> </span>Windell, Dylan<br class=""><b class="">Cc:</b><span class="apple-converted-space"> </span><a href="mailto:openspim@openspim.org" class="" style="color: purple; text-decoration: underline;"><span class="" style="color: purple;">openspim@openspim.org</span></a><br class=""><b class="">Subject:</b><span class="apple-converted-space"> </span>Re: [OpenSPIM] 3D Reconstruction</span></div></div></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">Dear Dylan,</div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><br class=""><br class=""><br class=""><br class=""></div></div></div><div class=""><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;">I have been trying to register Z-stacks of Zebrafish using the SPIM plugin with various concentrations of beads but despite everything I try I cannot get the beads to match with different angles.</span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;"> </span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;">I am not sure if this is because of the light sheet not being aligned properly (I am calculating the PSF very soon and will re-align the system from scratch) or due to sample prep etc. I have corrected bead drifting (as this was evident in low concentrations of agarose) and I am pretty sure the xy and z resolution numbers are correct. I will attempt the Multiview reconstruction package, FluoRender or even ClearVolume but without bead registration working I am not sure if that will work either.</span></div></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">Did you already run the multi-view reconstruction? Can you attach the ImageJ log. I am particularly looking for numbers of beads and numbers of RANSAC inliers. That would be the way to start troubleshooting.<br class=""><br class=""><br class=""><br class=""></div></div></div><div class=""><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;"> </span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;">Would it be possible that if the FEP tube is not straight, the manual positioning of the next angles would misalign bead registration? I have also thought that as the embryo fills up the tube, I am not getting enough volume of beads on either side of the stack for bead registration?</span></div></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">That maybe a problem. There may be not enough beads in the OVERLAP between the views. You can visualise that in the Multiview Reconstruction application.</div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">I will help more after you send the logs or even deposit the data somewhere.</div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">All the best</div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">PAvel</div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"> </div></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><br class=""><br class=""><br class=""><br class=""></div></div></div><div class=""><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;"> </span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;">If anyone has had difficulty in this or have done work on Zebrafish with SPIM I would really appreciate any help.</span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;"> </span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;">Kind regards,</span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;"> </span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;">Dylan</span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;"> </span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 10pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Dylan Windell</span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 10pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Postgraduate Researcher</span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 10pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);"><a href="http://business-school.exeter.ac.uk/research/areas/centres/isr/" class="" style="color: purple; text-decoration: underline;"><span class="" style="color: rgb(31, 73, 125);">College</span></a> of Life and Environmental Sciences</span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 10pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">University of Exeter, Geoffrey Pope</span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 10pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">Stocker Road, Exeter EX4 4QD, UK</span></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><i class=""><span class="" style="font-size: 10pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">phone: +44 (0) 7596725056</span></i></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><i class=""><span class="" style="font-size: 10pt; font-family: Calibri, sans-serif; color: rgb(31, 73, 125);">email: <u class=""><a href="mailto:dw388@exeter.ac.uk" class="" style="color: purple; text-decoration: underline;"><span class="" style="color: purple;">dw388@exeter.ac.uk</span></a></u></span></i></div></div></div></div><div class=""><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;"><span class="" style="font-size: 11pt; font-family: Calibri, sans-serif;"> </span></div></div></div></div><div class=""><div class=""><div class="" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: 'Times New Roman', serif;">_______________________________________________<br class="">OpenSPIM mailing list<br class=""><a href="mailto:OpenSPIM@openspim.org" class="" style="color: purple; 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