[OpenSPIM] 3D Reconstruction

[Windell, Dylan]

Hi All,

I have been trying to register Z-stacks of Zebrafish using the SPIM plugin with various concentrations of beads but despite everything I try I cannot get the beads to match with different angles.

I am not sure if this is because of the light sheet not being aligned properly (I am calculating the PSF very soon and will re-align the system from scratch) or due to sample prep etc. I have corrected bead drifting (as this was evident in low concentrations of agarose) and I am pretty sure the xy and z resolution numbers are correct. I will attempt the Multiview reconstruction package, FluoRender or even ClearVolume but without bead registration working I am not sure if that will work either.

Would it be possible that if the FEP tube is not straight, the manual positioning of the next angles would misalign bead registration? I have also thought that as the embryo fills up the tube, I am not getting enough volume of beads on either side of the stack for bead registration?

If anyone has had difficulty in this or have done work on Zebrafish with SPIM I would really appreciate any help.

Kind regards,

Dylan

Dylan Windell
Postgraduate Researcher
College<http://business-school.exeter.ac.uk/research/areas/centres/isr/> of Life and Environmental Sciences
University of Exeter, Geoffrey Pope
Stocker Road, Exeter EX4 4QD, UK
phone: +44 (0) 7596725056
email: dw388 at exeter.ac.uk<mailto:dw388 at exeter.ac.uk>

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