[OpenSPIM] Information on T-SPIM construction

[David Burk]

Thanks, Michael. And thanks, Carsten for your link to the T-SPIM chamber
picture.

Best,
David

On Sat, Feb 21, 2015 at 6:24 AM, Michael Weber <weber at mpi-cbg.de> wrote:

> Hi David,
>
> you are right, with "blurry" I was referring to those parts of an image
> that are illuminated with a scattered/absorbed/refracted light sheet. You
> can of course adjust your system in several ways, either by making the
> light sheets more homogeneous and using simultaneous illumination, or by
> placing the waists of two thin "left" and "right" light sheets next to each
> other and using sequential acquisitions. Something to keep in mind is that
> sequential acquisitions take at least twice as long, require twice as much
> space and you have to merge both sides in a post-processing step. My
> recommendation is, as you already suggested, to start with a simple system
> with a 50/50 splitter and manual shutters.
>
> Best,
> Michael
>
> On Feb 20, 2015, at 3:58 PM, David Burk <davidburk1972 at gmail.com> wrote:
>
> Thanks, Michael.
>
> Depending on the sample, this might not be ideal, as you get an overlay of
>> "sharp" and "blurry" images.
>
> Is this due to the fact that one light sheet, say from the left, may not
> have its minimum beam waist at the same position as that from that of the
> right sheet? Or more due to scattering, absorption, etc. of the light as it
> interacts with the specimen?
> I found some images that showed what you are talking about. I just thought
> there may be some 'middle ground' you could settle on by adjusting the
> sheets from either side that would give relatively uniform illumination
> (but with a loss of resolution).
> It does sound like - if you choose the slow manual method - this could be
> doable. Illuminate from left > capture images > close left shutter >
> illuminate from right > capture > merge?
> Perhaps detailed designs of a T-SPIM system will eventually be uploaded to
> the web along with the modifications to the software to drive the
> additional hardware.
>
> Have a great weekend,
> David
>
> On Fri, Feb 20, 2015 at 6:40 AM, Michael Weber <weber at mpi-cbg.de> wrote:
>
>> Hey David,
>>
>> Pete might have a step file for the 3-objective chamber. And yes, in
>> principle you just need two arms and a 50/50 splitter, but there are some
>> limitations. Unless you add shutters or AOTFs, you simultaneously
>> illuminate from both sides. Depending on the sample, this might not be
>> ideal, as you get an overlay of "sharp" and "blurry" images. This typically
>> gets worse with larger samples. You can add manual shutters to be able to
>> choose the side of illumination. If you want to be able to quickly record
>> images with sequential double-sided illumination, you need to integrate
>> shutters/AOTFs into the software.
>>
>> Best,
>> Michael
>>
>> On Feb 18, 2015, at 8:41 PM, David Burk <davidburk1972 at gmail.com> wrote:
>>
>> I apologize in advance if the answer to my question is somewhere in the
>> archives.
>>
>> I am interested in building a T-SPIM system and ran across an archived
>> conversation on this from June 2014. There was mention that such a setup
>> had been built as well as a possible 3D model of the imaging chamber being
>> available. I couldn't find any additional information on the OpenSPIM.org
>> <http://openspim.org/> site and just wanted to ask if there was a link
>> to this information somewhere. Based on the 3D model on the site it looks
>> like you really just need to replicate one arm of the laser path and add a
>> 50/50 splitter near the laser. Would there be needed changes to the
>> operating software? If so, that is beyond my (rather limited) capabilities.
>> However, if instructions exist that'd be great!
>>
>> I appreciate any information you may have on this.
>>
>> Best,
>> David
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>>
>>
>>
>> _____________
>
> 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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