<html><head><meta http-equiv="Content-Type" content="text/html; charset=utf-8"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">Dear Alessandro<div class=""><br class=""></div><div class="">We have imaged pupae held with glass capillaries, where they have air supply from the top. I guess it does not matter if the capillary is held from the top or bottom as long as it is open on the top. You can also try FEP tubes.</div><div class=""><br class=""></div><div class=""><a href="https://www.ncbi.nlm.nih.gov/pubmed/19465594" class="">https://www.ncbi.nlm.nih.gov/pubmed/19465594</a></div><div class=""><br class=""></div><div class=""><img apple-inline="yes" id="9DC7BDEE-E1F4-4FB1-A596-256F7E87FA56" width="243" height="368" src="cid:143B93C2-6020-4BAB-956C-2FC13612E416@attlocal.net" class=""></div><div class=""><br class=""></div><div class="">Best</div><div class="">Jan<br class=""><div><br class=""><blockquote type="cite" class=""><div class="">On Apr 12, 2018, at 5:02 AM, Alessandro Ciccarelli <<a href="mailto:alessandro.ciccarelli@crick.ac.uk" class="">alessandro.ciccarelli@crick.ac.uk</a>> wrote:</div><br class="Apple-interchange-newline"><div class=""><div class="WordSection1" style="page: WordSection1; font-family: Helvetica; font-size: 12px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px;"><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">Dear light-sheet users,<o:p class=""></o:p></span></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">does anybody have experience in long-term imaging of drosophila pupae with light-sheet? I`m currently looking for suggestions about how to mount this tricky sample; actually the pupa needs to be:<o:p class=""></o:p></span></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">- partially in contact with air (it needs to breath)<o:p class=""></o:p></span></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">- immersed in water (I`m using water dipping lenses)<o:p class=""></o:p></span></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""> </o:p></span></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">I`m currently using a Luxendo MuVi-SPIM, where the holder is coming from the bottom of the chamber.<o:p class=""></o:p></span></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">Thanks in advance!<o:p class=""></o:p></span></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">Best,<o:p class=""></o:p></span></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">Alessandro<o:p class=""></o:p></span></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""> </o:p></span></div><div id="Signature" class=""><div id="divtagdefaultwrapper" class=""><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="normaltextrun"><span lang="EN-US" style="color: rgb(127, 127, 127);" class="">---</span></span><span class="eop"><span lang="EN-US" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="normaltextrun"><span lang="EN-US" style="color: rgb(127, 127, 127);" class="">Alessandro Ciccarelli </span></span><span class="eop"><span lang="EN-US" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="normaltextrun"><i class=""><span lang="EN-US" style="color: rgb(127, 127, 127);" class="">Light Sheet Specialist</span></i></span><span class="eop"><span lang="EN-US" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="normaltextrun"><span lang="EN-US" style="color: rgb(127, 127, 127);" class="">CALM - Crick Advanced Light Microscopy Facility</span></span><span class="eop"><span lang="EN-US" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="normaltextrun"><span lang="EN-US" style="color: rgb(127, 127, 127);" class="">The Francis Crick Institute</span></span><span class="eop"><span lang="EN-US" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="normaltextrun"><span lang="EN-US" style="color: rgb(127, 127, 127);" class="">1 Midland Road - London -NW1 1AT</span></span><span class="eop"><span lang="EN-US" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="normaltextrun"><span lang="EN-US" style="color: rgb(127, 127, 127);" class="">United Kingdom</span></span><span class="eop"><span lang="EN-US" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="eop"><span lang="EN-US" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="normaltextrun"><span lang="EN-US" style="color: rgb(127, 127, 127);" class="">T: +442037960913</span></span><span class="eop"><span lang="EN-US" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="normaltextrun"><span lang="EN-US" style="color: rgb(127, 127, 127);" class="">E: </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><a href="mailto:alessandro.ciccarelli@crick.ac.uk" target="_blank" title="mailto:alessandro.ciccarelli@crick.ac.uk Ctrl+Click or tap to follow the link" id="LPNoLP" style="color: purple; text-decoration: underline;" class=""><span class="normaltextrun"><span style="font-family: "Times New Roman", serif; color: rgb(127, 127, 127);" class="">alessandro.ciccarelli@crick.ac.uk</span></span></a></span><span class="eop"><span lang="EN-US" style="font-size: 11pt;" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; vertical-align: baseline;" class=""><span class="normaltextrun"><span lang="EN-US" style="color: rgb(127, 127, 127);" class="">W: </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><a href="http://www.crick.ac.uk/" target="_blank" id="LPNoLP" style="color: purple; text-decoration: underline;" class=""><span class="normaltextrun"><span style="font-family: "Times New Roman", serif; color: rgb(127, 127, 127);" class="">www.crick.ac.uk</span></span></a></span><span class="eop"><span lang="EN-US" style="font-size: 11pt; font-family: Arial, sans-serif;" class=""> </span></span><span lang="EN-US" style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""> </o:p></span></div></div></div><p class="MsoNormal" style="margin: 0cm 0cm 12pt; font-size: 12pt; font-family: "Times New Roman", serif;"><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""> </o:p></span></p><div class=""><div class="MsoNormal" align="center" style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif; text-align: center;"><span style="font-family: Calibri, sans-serif;" class=""><hr size="2" width="98%" align="center" class=""></span></div><div id="divRplyFwdMsg" class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><b class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class="">From:</span></b><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""><span class="Apple-converted-space"> </span>OpenSPIM <<a href="mailto:openspim-bounces@openspim.org" style="color: purple; text-decoration: underline;" class="">openspim-bounces@openspim.org</a>> on behalf of Jan Huisken <<a href="mailto:huisken@mpi-cbg.de" style="color: purple; text-decoration: underline;" class="">huisken@mpi-cbg.de</a>><br class=""><b class="">Sent:</b><span class="Apple-converted-space"> </span>09 April 2018 21:52<br class=""><b class="">To:</b><span class="Apple-converted-space"> </span>Feinstein, Timothy N;<span class="Apple-converted-space"> </span><a href="mailto:openspim@openspim.org" style="color: purple; text-decoration: underline;" class="">openspim@openspim.org</a><br class=""><b class="">Subject:</b><span class="Apple-converted-space"> </span>Re: [OpenSPIM] holographic diffusers</span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class=""> <o:p class=""></o:p></span></div></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">Hi Tim<o:p class=""></o:p></span></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""> </o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">We have not yet tried those diffusers.<o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""> </o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">Why is your mSPIM not working? Sounds like your resonant mirror is not in the right plane. Please let us know if there is anything we can help with.<o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""> </o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">Best<o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">Jan<o:p class=""></o:p></span></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class=""><br class=""><br class=""><o:p class=""></o:p></span></div><blockquote style="margin-top: 5pt; margin-bottom: 5pt;" class=""><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class="">On Apr 9, 2018, at 8:50 AM, Feinstein, Timothy N <<a href="mailto:tnf8@PITT.EDU" id="LPlnk578761" style="color: purple; text-decoration: underline;" class="">tnf8@PITT.EDU</a>> wrote:<o:p class=""></o:p></span></div></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""> </o:p></span></div><div class=""><div class=""><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class="">Hi folks, </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""> </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class="">Salili and co-workers (Review Sci. Instruments 89: 036107) just published a very interesting technique for eliminating stripe artifacts using elliptical holographic diffusers. It looks like a much simpler version of mSPIM, with just a couple filters and no moving parts. This would be great news for me since my mSPIM design doesn’t work – instead of tilting, the sheet moves up and down in a parallel track. </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""> </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class="">Has anyone tried holographic diffusers before? They look like they need to be quite close to the sample, so it’s possible they would not work with a conventional openSPIM sample holder. Any feedback would be appreciated. </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""> </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class="">Thanks!</span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""> </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""><br class="">Tim</span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""> </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10.5pt; font-family: Calibri, sans-serif;" class="">Timothy Feinstein, Ph.D. </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10.5pt; font-family: Calibri, sans-serif;" class="">Research Scientist</span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10.5pt; font-family: Calibri, sans-serif;" class="">Department of Developmental Biology</span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 10.5pt; font-family: Calibri, sans-serif;" class="">University of Pittsburgh</span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""> </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div><div class=""><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 11pt; font-family: Calibri, sans-serif;" class=""> </span><span style="font-family: Calibri, sans-serif;" class=""><o:p class=""></o:p></span></div></div></div><div style="margin: 0cm 0cm 0.0001pt; font-size: 12pt; font-family: "Times New Roman", serif;" class=""><span style="font-size: 9pt; font-family: Helvetica, sans-serif;" class="">_______________________________________________<br class="">OpenSPIM mailing list<br class=""></span><span style="font-family: Calibri, sans-serif;" class=""><a href="mailto:OpenSPIM@openspim.org" id="LPlnk662653" style="color: purple; text-decoration: underline;" class=""><span style="font-size: 9pt; font-family: Helvetica, sans-serif; color: rgb(149, 79, 114);" class="">OpenSPIM@openspim.org</span></a></span><span style="font-size: 9pt; font-family: Helvetica, sans-serif;" class=""><br class=""></span><span style="font-family: Calibri, sans-serif;" class=""><a href="http://openspim.org/mailman/listinfo/openspim" id="LPlnk459532" style="color: purple; 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