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Gabriella Turek
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One of the reasons for running the OpenSPIM project is to provide a
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platform for teaching light-sheet microscopy. At the end of March 2013
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we have brought OpenSPIM to an EMBO course in South Africa. Here are
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[Pavel
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Tomancak](http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html)'s
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first hand impressions on how it worked out: ![Imaging Infection and
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Immunity](SA_course_poster.jpg "Imaging Infection and Immunity")
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One of the reasons for running the OpenSPIM project is to provide a platform for teaching light-sheet microscopy. At the end of March 2013 we have brought OpenSPIM to an EMBO course in South Africa. Here are [Pavel Tomancak](https://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html)'s first hand impressions on how it worked out:
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The EMBO course [Imaging Infection and
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Immunity](http://microscopy.synbio.scientific-solution.com/) is a
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practical course aimed at researchers studying infectious diseases with
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strong emphasis on imaging. It has been organized three times over the
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last 4 years by **Musa Mhlanga, Freddy Frischknecht and Jost Enninga**.
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The first installment of the course has been held at the
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[CSIR](http://www.csir.co.za/) in Pretoria South Africa right before the
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World Cup and it stayed there ever since. ![Freddy Frischknecht, Musa
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Mhlanga, Jost Enninga](Freddy_Musa_Jost.jpg
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"Freddy Frischknecht, Musa Mhlanga, Jost Enninga")
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{% include image src="SA_course_poster.jpg" width="70%" caption="Imaging Infection and Immunity" %}
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Pete Pitrone attended the entire course. He managed to get the OpenSPIM
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in a suitcase onto an airplane and with the necessary paperwork in hand
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passed it through the customs without any problems. This is good news
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since it indicates that we can actually travel to a foreign country by
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plane bringing a set-up that admittedly looks like a bomb. Freddy
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Frischknecht also brought parts for his OpenSPIM. Unfortunately he was
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missing the most important pieces - the objectives - and so there was no
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imaging on what I called [on Twitter](https://twitter.com/PavelTomancak)
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*OpenSPIM's little Heidelberg bro* (the slang was necessary due to
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The EMBO course [Imaging Infection and Immunity](https://microscopy.synbio.scientific-solution.com/) is a practical course aimed at researchers studying infectious diseases with strong emphasis on imaging. It has been organized three times over the last 4 years by **Musa Mhlanga, Freddy Frischknecht and Jost Enninga**. The first installment of the course has been held at the [CSIR](https://www.csir.co.za/) in Pretoria South Africa right before the World Cup and it stayed there ever since. ![Freddy Frischknecht, Musa Mhlanga, Jost Enninga](Freddy_Musa_Jost.jpg "Freddy Frischknecht, Musa Mhlanga, Jost Enninga")
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Pete Pitrone attended the entire course. He managed to get the OpenSPIM in a suitcase onto an airplane and with the necessary paperwork in hand passed it through the customs without any problems. This is good news since it indicates that we can actually travel to a foreign country by plane bringing a set-up that admittedly looks like a bomb. Freddy Frischknecht also brought parts for his OpenSPIM. Unfortunately he was missing the most important pieces - the objectives - and so there was no imaging on what I called [on Twitter](https://twitter.com/PavelTomancak) *OpenSPIM's little Heidelberg bro* (the slang was necessary due to
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Twitter character limits).
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Pete built the system four times with the students during the course. We
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wanted to image His-YFP expressing *Drosophila* however it was
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relatively difficult to prepare the *Drosophila* samples in Musa's lab
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which works mostly with mammalian cells. Some crucial pieces of
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equipment were missing - like paint brush and sieves and so until I
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arrived the students managed to image only some random critters picked
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up from the local pond. I was able to later on improvise the
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*Drosophila* sample prep (the sieves can be substituted reasonably by
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*two* lens cleaning tissues and instead of the brush one can use a
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feather plucked from some random African bird ;-)). ![Pete Pitrone
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explaining SPIM principles to ALA students](Pete_ALA.jpg
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"Pete Pitrone explaining SPIM principles to ALA students")
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Pete built the system four times with the students during the course. We wanted to image His-YFP expressing *Drosophila* however it was relatively difficult to prepare the *Drosophila* samples in Musa's lab which works mostly with mammalian cells. Some crucial pieces of equipment were missing - like paint brush and sieves and so until I arrived the students managed to image only some random critters picked up from the local pond. I was able to later on improvise the *Drosophila* sample prep (the sieves can be substituted reasonably by *two* lens cleaning tissues and instead of the brush one can use a feather plucked from some random African bird ;-)).
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{% include image src="Pete_ALA.jpg" width="70%" caption="Pete Pitrone explaining SPIM principles to ALA students" %}
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During the course we discovered a problem with the software - the
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Micro-Manager plugin refused to save anything. Fortunately the good
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people in Madison, specifically Johannes Schindelin, rescued us as they
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always do by providing a hot fix for the software issue.
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During the course we discovered a problem with the software - the Micro-Manager plugin refused to save anything. Fortunately the good people in Madison, specifically Johannes Schindelin, rescued us as they always do by providing a hot fix for the software issue.
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It happened just in time, because the next day the course was visited by
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high school students from the **African Leadership Academy**
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[ALA](http://www.africanleadershipacademy.org/). I presented a talk
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tailored for younger, non-scientific audience centered around SPIM
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imaging. It actually wasn't necessary to dumb things down, since many of
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the students had strong biological background and all, even the particle
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physicists (;-)), were very smart. Pete and Jessica from Freddy's lab
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built the set-up with them. This event attracted serious media
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attention. South African TV was shooting a documentary about women in
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science and they got so intrigued by our do-it-yourself microscope that
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they will include it. The ALA students were giving professional
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interviews. Musa, myself and Pete also talked some non-sense into the
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camera. I dread what will come out of that, but overall this was a great
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outreach event. ![Pavel and Pete 'posing' with OpenSPIM in a
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suitcase](Pavel_Pete_OpenSPIM.jpg
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"Pavel and Pete 'posing' with OpenSPIM in a suitcase")
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It happened just in time, because the next day the course was visited by high school students from the **African Leadership Academy** [ALA](https://www.africanleadershipacademy.org/). I presented a talk tailored for younger, non-scientific audience centered around SPIM imaging. It actually wasn't necessary to dumb things down, since many of the students had strong biological background and all, even the particle physicists (;-)), were very smart. Pete and Jessica from Freddy's lab built the set-up with them. This event attracted serious media attention. South African TV was shooting a documentary about women in science and they got so intrigued by our do-it-yourself microscope that they will include it. The ALA students were giving professional interviews. Musa, myself and Pete also talked some non-sense into the camera. I dread what will come out of that, but overall this was a great outreach event.
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The feedback on the wiki from the course students was overwhelmingly
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positive. In their final report they called it *absolutely idiot proof*.
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We observed what resources they were using when building the set-up and
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found out that they focus mainly on the static SolidWorks and real
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pictures - building it like a Lego. My impression was that especially
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the students from Africa highly appreciate the fact that we are making
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this technology accessible.
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{% include image src="Pavel_Pete_OpenSPIM.jpg" width="70%" caption="Pavel and Pete 'posing' with OpenSPIM in a suitcase" %}
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We managed on the last day of the course to record a short but almost
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perfect time-lapse of His-YFP expressing *Drosophila* embryo from 6
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angles. I will call that a success, we brought down to South Africa bits
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and pieces and we managed to put together a completely functional SPIM
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set-up.
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The feedback on the wiki from the course students was overwhelmingly positive. In their final report they called it *absolutely idiot proof*. We observed what resources they were using when building the set-up and found out that they focus mainly on the static SolidWorks and real pictures - building it like a Lego. My impression was that especially the students from Africa highly appreciate the fact that we are making this technology accessible. We managed on the last day of the course to record a short but almost perfect time-lapse of His-YFP expressing *Drosophila* embryo from 6 angles. I will call that a success, we brought down to South Africa bits and pieces and we managed to put together a completely functional SPIM set-up.
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We encountered a lot of problems and learned some valuable lessons.
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- don't underestimate the sample prep, bring absolutely everything
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- if possible bring a big, fast computer for the image processing
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- ask everyone to download Fiji before the course even starts
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- test the set-up before you go
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- tell everyone that the course is going to happen - especially
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Madison - so that they can brace for emergencies
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- bring protective goggles for the students to have at least basic
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laser safety (as far as I know nobody got blinded during the
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course).
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- design a box to cover the sample chamber - likely you won't image in
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a microscopy room and Africa is - well - bright
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- tell everyone that the course is going to happen - especially Madison - so that they can brace for emergencies
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- bring protective goggles for the students to have at least basic laser safety (as far as I know nobody got blinded during the course).
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- design a box to cover the sample chamber - likely you won't image in a microscopy room and Africa is - well - bright
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The next course that will feature OpenSPIM will be the EMBO practical
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course on [Marine animal models in evolution &
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development](http://events.embo.org/13-marine-devo/index.html) in Sweden
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in July.
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The next course that will feature OpenSPIM will be the EMBO practical course on [Marine animal models in evolution & development](https://events.embo.org/13-marine-devo/index.html) in Sweden in July.
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PAvel
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Pavel
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[Pavel
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Tomancak](http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html)
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contributed a [detailed
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tutorial](http://fiji.sc/SPIM_Registration_on_cluster) how to analyze
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SPIM images on a cluster using Fiji.
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[Pavel Tomancak](httpS://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html) contributed a [detailed tutorial](https://fiji.sc/SPIM_Registration_on_cluster) how to analyze SPIM images on a cluster using Fiji.
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[OpenSPIM](http://www.nature.com/nmeth/journal/vaop/ncurrent/full/nmeth.2507.html)
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was published in Nature Methods, back to back with a paper on a similar
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project - the
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[OpenSpinMicroscopy](http://www.nature.com/nmeth/journal/vaop/ncurrent/full/nmeth.2508.html).
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[OpenSPIM](https://www.nature.com/nmeth/journal/vaop/ncurrent/full/nmeth.2507.html) was published in Nature Methods, back to back with a paper on a similar project - the [OpenSpinMicroscopy](https://www.nature.com/nmeth/journal/vaop/ncurrent/full/nmeth.2508.html).
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For more details see also our much longer pre-print [on
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arXiv](http://arxiv.org/abs/1302.1987).
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For more details see also our much longer pre-print [on arXiv](https://arxiv.org/abs/1302.1987).
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After you build an OpenSPIM and do something cool with it please
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remember to cite us\!
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After you build an OpenSPIM and do something cool with it please remember to cite us!
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Pitrone P. G., Schindelin J., Stuyvenberg L., Preibisch S., Weber M.;
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Eliceiri K. W., Huisken J., Tomancak P. (2013) **OpenSPIM: an open
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access light sheet microscopy platform** *Nat. Methods* in press
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[PDF](Media:Nmeth.2507.pdf "wikilink")
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[Supplement](Media:Nmeth.2507-S1.pdf "wikilink")
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Pitrone P. G., Schindelin J., Stuyvenberg L., Preibisch S., Weber M.; Eliceiri K. W., Huisken J., Tomancak P. (2013) **OpenSPIM: an open access light sheet microscopy platform** *Nat. Methods* in press [PDF](documents/Nmeth.2507.pdf) [Supplement](document/Nmeth.2507-S1.pdf)
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In July 2013 we brought the OpenSPIM to a EMBO course on
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\[<http://events.embo.org/13-marine-devo/>|**Marine animal models in
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evolution & development**\] (MAMED) at Sven Lovén Centre for Marine
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Sciences in Kristineberg in Sweden. The course is an annual event
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organized primarily by Detlev Arendt, this year together with Andreas
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Hejnoj and Grahame Budd. The course features a vast array of amazing
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speakers focusing on development of marine model organisms such as
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Platynereis, sea urchin, parhyale, flatworms, ciona and many other
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species representing different animal phyla many of which you have never
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heard about before. There are really interesting talks introducing the
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different animal models and the cutting edge science done with them.
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There are a lot of discussions about evolution of animal diversity
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driven by the leaders of the field. Since this is a practical course the
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emphasis is on doing experiments which involves collecting the animals
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on rather adventurous boat trips around the fjord which apparently
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features representatives of most phyla. The collected samples are sifted
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through to find interesting specimen, typically identified by Claus
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Nielsen who knows simply everything about them. They are then fixed,
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stained by RNA i*n situ* or antibody staining or microinjected for live
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imaging.
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In July 2013 we brought the OpenSPIM to a EMBO course on [**Marine animal models in evolution & development**](https://events.embo.org/13-marine-devo/) at Sven Lovén Centre for Marine Sciences in Kristineberg in Sweden. The course is an annual event organized primarily by Detlev Arendt, this year together with Andreas Hejnoj and Grahame Budd. The course features a vast array of amazing speakers focusing on development of marine model organisms such as Platynereis, sea urchin, parhyale, flatworms, ciona and many other species representing different animal phyla many of which you have never heard about before. There are really interesting talks introducing the different animal models and the cutting edge science done with them. There are a lot of discussions about evolution of animal diversity driven by the leaders of the field. Since this is a practical course the emphasis is on doing experiments which involves collecting the animals on rather adventurous boat trips around the fjord which apparently features representatives of most phyla. The collected samples are sifted through to find interesting specimen, typically identified by Claus Nielsen who knows simply everything about them. They are then fixed, stained by RNA i*n situ* or antibody staining or microinjected for live imaging.
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Which is why we were there. The course this year focussed on light sheet
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microscopy and next to OpenSPIM there was also
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\[<http://microscopy.zeiss.com/microscopy/en_de/products/imaging-systems/lightsheet-z-1.html>|**Lightsheet
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Z.1 from Carl Zeiss Microimaging**\].
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Which is why we were there. The course this year focussed on light sheet microscopy and next to OpenSPIM there was also [**Lightsheet Z.1 from Carl Zeiss Microimaging**](https://microscopy.zeiss.com/microscopy/en_de/products/imaging-systems/lightsheet-z-1.html)
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The Human Frontier Science Program has [written a success story about
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us](http://www.hfsp.org/frontier-science/hfsp-success-stories/openspim-open-access-light-sheet-microscopy-platform)\!
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The Human Frontier Science Program has [written a success story about us](https://www.hfsp.org/frontier-science/hfsp-success-stories/openspim-open-access-light-sheet-microscopy-platform)!
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Michael Weber from Jan Huisken lab has put together a nice protocol page
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describing [preparation of zebrafish embryos for multi-view SPIM
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imaging](Zebrafish_embryo_sample_preparation "wikilink"). Lots of
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photographs\!
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Michael Weber from Jan Huisken lab has put together a nice protocol page describing [preparation of zebrafish embryos for multi-view SPIM imaging](Zebrafish_embryo_sample_preparation). Lots of photographs!
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For more information see the JoVE video of the protocol
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- [Multilayer Mounting for Long-term Light Sheet Microscopy of
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Zebrafish](http://www.jove.com/video/51119/multilayer-mounting-for-long-term-light-sheet-microscopy-of-zebrafish)
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- [Multilayer Mounting for Long-term Light Sheet Microscopy of Zebrafish](https://www.jove.com/video/51119/multilayer-mounting-for-long-term-light-sheet-microscopy-of-zebrafish)
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and the paper in Development
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- [Multilayer mounting enables long-term imaging of zebrafish
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development in a light sheet
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microscope](http://dev.biologists.org/content/139/17/3242.abstract)
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- [Multilayer mounting enables long-term imaging of zebrafish development in a light sheet microscope](https://dev.biologists.org/content/139/17/3242.abstract)

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