[OpenSPIM] Imaging drosophila pupae

[Alessandro Ciccarelli]

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> on behalf of Jan Huisken <huisken at mpi-cbg.de>
Sent: 09 April 2018 21:52
To: Feinstein, Timothy N; 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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