[OpenSPIM] OpenSPIM and optically cleared 3D tissue

[Michael Weber]

Hi Jamie,

if you are using hazardous clearing media and don't have a dedicated dipping objective available, one idea would be to use long-working distance air objectives and a smaller, closed glass chamber. The main issue I would expect from such a combination would be a drift of your detection focal plane when moving the sample through the light sheet, because the light has to travel through layers of different refractive indices, and you change the "thickness" of those layers. But this drift should be linear and could be corrected by adapting the distance between light sheet and detection objective while acquiring a z-stack. So you would either need to motorize the mirror for translating the light sheet, or your detection objective, and then implement the drift correction in the software. Might be worth a try, unless I overlooked something important here.

Cheers,
Michael

On Oct 24, 2015, at 8:27 AM, Jamie Flynn <Jamie.Flynn at uon.edu.au> wrote:

> Hi Pete,
> 
> Thanks for the link. Some very impressive images from the mesolens. You¹re
> right that pickings are slim for commercially available lenses. Leica and
> Olympus have some objectives that are suitable for various types of
> cleared tissue, but come with hefty price tags. Apparently Zeiss have
> something in the works as well (not that it¹ll be cheap!). The RI range of
> imaging media for different clearing techniques is obviously one of the
> bigger challenges (e.g. 1.45 for Focusclear and 1.56 for DBE). Will look
> into a custom design as you suggest.
> 
> Cheers
> Jamie
> 
> 
> 
> On 23/10/2015 7:16 pm, "Peter Gabriel Pitrone" <pitrone at mpi-cbg.de> wrote:
> 
>> with the exeption of the "Meso-lens" from Prof. Brad Amos... But I'm sure
>> it uses glue, and you probably can not afford it.
>> 
>> http://www2.mrc-lmb.cam.ac.uk/research/technology-transfer/mesolens-micros
>> copy/
>> 
>> 
>> 
>> ----- Original Message -----
>> From: "Peter Gabriel Pitrone" <pitrone at mpi-cbg.de>
>> To: "Jamie Flynn" <Jamie.Flynn at uon.edu.au>
>> Cc: openspim at openspim.org
>> Sent: Friday, October 23, 2015 10:14:06 AM
>> Subject: Re: [OpenSPIM] OpenSPIM and optically cleared 3D tissue
>> 
>> Hey Jamie,
>> 
>> I would have someone design the lenses (and therefor chamber as well) for
>> you made in a way the is fluid/liquid proof WITHOUT using glue, also
>> keeping in mind the absolute largest (NA in terms of detection lens, and)
>> working distance possible so you could put in bigger tissues. I doubt
>> that there is a objective on the market that would meet your requirements.
>> 
>> Regards,
>> Pete
>> 
>> ----- Original Message -----
>> From: "Jamie Flynn" <Jamie.Flynn at uon.edu.au>
>> To: openspim at openspim.org
>> Sent: Friday, October 23, 2015 1:58:29 AM
>> Subject: [OpenSPIM] OpenSPIM and optically cleared 3D tissue
>> 
>> Hi everyone, 
>> 
>> We¹re building a dual illumination OpenSPIM at the Hunter Medical
>> Research Institute in Newcastle, Australia. Our ai m is to have the scope
>> and analysis pipeline used as a core facility for lab groups to image 3D
>> tumour cell cultures, developing embryos and most importantly for us,
>> optically cleared 3D tissue samples (e.g. CLARITY, CUBIC, 3DISCO, Scale
>> etc.). Imaging cleared 3D samples would be an interesting avenue for
>> OpenSPIM and it¹d be great to hear about any hardware modifications or
>> mounting protocols others may have used to do it.
>> 
>> If anyone wants to try clearing out their own tissue samples, my
>> colleagues and I put together a handbook that describes the reagents,
>> equipment and detailed protocols for ŒPassive CLARITY¹. We compiled the
>> protocols outlined in Tomer et al. Nat. Prot. 9(7) 2014 and Yang et al.
>> Cell (158) 2014 and added in our own refinements (after much trial and
>> error!). It¹s a very simple technique and has worked really well in our
>> hands. Here¹s the link. Download it to your desktop in case the link gets
>> broken: 
>> 
>> https://drive.google.com/open?id=0BzIZDgZFR2srNFk2dHFqT1UycFk
>> 
>> All the best 
>> --------------------
>> Jamie Flynn (PhD) 
>> University of Newcastle
>> Hunter Medical Research Institute
>> Room 3613 level 3 east
>> 1 Kookaburra Circuit, New Lambton Heights
>> NSW 2305, Australia.
>> pH: +612 40420468 
>> Mobile: +61 416003787
>> Email: jamie.flynn at uon.edu.au

_____________

Michael Weber
Postdoc, Huisken lab
Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108, 01307 Dresden
Tel. 0049 351/2102837

http://www.mpi-cbg.de/huisken

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