[OpenSPIM] Multiview registration - no inliers
Stephan.Preibisch at mdc-berlin.de
Stephan.Preibisch at mdc-berlin.de
Fri Aug 19 10:58:48 CDT 2016
Hi, my first guess would be that your calibration (pixel size is x,y and z) is wrong ... Can you make sure this is correct when you define your dataset?
All the best,
Stephan
On Aug 19, 2016, at 17:17, Tania Mendonca <tvmendonca1 at sheffield.ac.uk<mailto:tvmendonca1 at sheffield.ac.uk>> wrote:
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 detecting 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
--
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Tania Mendonca
Doctoral Research Student
University of Sheffield
S10 2TN
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