[OpenSPIM] Imaging drosophila pupae

[Jan Huisken]

Dear Alessandro

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.

https://www.ncbi.nlm.nih.gov/pubmed/19465594 <https://www.ncbi.nlm.nih.gov/pubmed/19465594>



Best
Jan

> On Apr 12, 2018, at 5:02 AM, Alessandro Ciccarelli <alessandro.ciccarelli at crick.ac.uk> wrote:
> 
> Dear light-sheet users,
> 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:
> - partially in contact with air (it needs to breath)
> - immersed in water (I`m using water dipping lenses)
>  
> I`m currently using a Luxendo MuVi-SPIM, where the holder is coming from the bottom of the chamber.
> Thanks in advance!
> Best,
> Alessandro
>  
> --- 
> Alessandro Ciccarelli  
> Light Sheet Specialist 
> CALM - Crick Advanced Light Microscopy Facility 
> The Francis Crick Institute 
> 1 Midland Road - London -NW1 1AT 
> United Kingdom 
>  
> T: +442037960913 
> E: alessandro.ciccarelli at crick.ac.uk <mailto:alessandro.ciccarelli at crick.ac.uk> 
> W: www.crick.ac.uk <http://www.crick.ac.uk/> 
>  
>  
> 
> From: OpenSPIM <openspim-bounces at openspim.org <mailto:openspim-bounces at openspim.org>> on behalf of Jan Huisken <huisken at mpi-cbg.de <mailto:huisken at mpi-cbg.de>>
> Sent: 09 April 2018 21:52
> To: Feinstein, Timothy N; openspim at openspim.org <mailto:openspim at openspim.org>
> Subject: Re: [OpenSPIM] holographic diffusers
>  
> Hi Tim
>  
> We have not yet tried those diffusers.
>  
> 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.
>  
> Best
> Jan
> 
> 
> On Apr 9, 2018, at 8:50 AM, Feinstein, Timothy N <tnf8 at PITT.EDU <mailto:tnf8 at PITT.EDU>> wrote:
>  
> Hi folks, 
>  
> 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. 
>  
> 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.  
>  
> Thanks!
>  
> 
> Tim
>  
> Timothy Feinstein, Ph.D. 
> Research Scientist
> Department of Developmental Biology
> University of Pittsburgh
>  
>  
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