[OpenSPIM] Multiview registration - no inliers

Tania Mendonca tvmendonca1 at sheffield.ac.uk
Fri Aug 19 10:16:01 CDT 2016


Hi all

Hope your microscopes are aligned and treating you well! I'm sorry if I've
missed this conversation before but I was wondering if any of you have had
similar problems as me with trying to get the Fiji multiview reconstruction
plugin to like my data.

I've tried a range of microspheres - different colours, different sizes and
different concentrations but it never seems to find enough correspondences
between views. I've been primarily experimenting with interactively
detec​​ting interest points. I've tried the presets as well but with no
luck. I've tried to systematically work through various combinations of
sigma and threshold but always end up with 0 inliers. Registrations always
terminates with the following error: "There are no connected tiles, cannot
do an optimization. Quitting."

My home built system doesn't run on µManager and my images are acquired as
.cxd files which I convert to .tifs before processing. I'm attaching some
of my test data - https://drive.google.com/file/d/0B-he-_wcys_-
aUtRZ0pidFhyUlk/view?usp=sharing
the sample here is 1µm yellow-green beads (1:4000 dilution) and 10µm beads
(1:50 dilution) with the idea of using the 1µm beads to reconstruct the
10µm beads. I use the same objectives as on the OpenSPIM system (10X for
illumination and 20X for detection), magnification on the system is 27.7X.
The illumination here is 473nm and detection at 520nm.​​

A log file has been included in the zip file to give you an idea of what
I've been trying.

I hope I'm just missing a trick somewhere. Any help would be greatly
appreciated!

Best wishes
Tania
-- 
-------------------------
Tania Mendonca
Doctoral Research Student
University of Sheffield
S10 2TN
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