<div dir="ltr">Hi<div><br></div><div>I've just checked to make sure but yes I have corrected the voxel size on the image stacks and there shouldn't be a mistake in that. However, your reply gave me an idea and I did a quick test. I duplicated one of the images and ran the multiview reconstruction plugin for the two duplicates. It worked, so it makes me think that there's nothing wrong with the files themselves and you might be right about the issue being with my voxel depth. I've referred to SVI's Nyquist calculator to workout my step size, is there a recommended step size that people use?</div><div><br></div><div>Best wishes</div><div>Tania</div></div><div class="gmail_extra"><br><div class="gmail_quote">On 19 August 2016 at 16:58, <a href="mailto:Stephan.Preibisch@mdc-berlin.de">Stephan.Preibisch@mdc-berlin.de</a> <span dir="ltr"><<a href="mailto:Stephan.Preibisch@mdc-berlin.de" target="_blank">Stephan.Preibisch@mdc-berlin.de</a>></span> wrote:<br><blockquote class="gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex">
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<div>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?</div>
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<div>All the best,</div>
<div>Stephan <br>
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On Aug 19, 2016, at 17:17, Tania Mendonca <<a href="mailto:tvmendonca1@sheffield.ac.uk" target="_blank">tvmendonca1@sheffield.ac.uk</a>> wrote:<br>
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<div dir="ltr">Hi all
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<div>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. </div>
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<div>I've tried a range of microspheres - different colours, different sizes and different concentrations but it never seems to find enough correspondences between views. <span style="font-size:12.8px">I</span><span style="font-size:12.8px">'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: </span><span style="font-size:12.8px">"There are no connected tiles, cannot do an optimization. Quitting."</span></div>
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<div>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 -
<a href="https://drive.google.com/file/d/0B-he-_wcys_-aUtRZ0pidFhyUlk/view?usp=sharing" target="_blank">
https://drive.google.com/file/<wbr>d/0B-he-_wcys_-aUtRZ0pidFhyUlk<wbr>/view?usp=sharing</a></div>
<div>the sample here is 1µm yellow-green beads <span style="font-size:12.8px">(1:4000 dilution) and 10µm beads (1:50 dilution) with the idea of using the 1µm beads to reconstruct the 10µm beads. </span><span style="font-size:12.8px">I use the same objectives
as on the OpenSPIM system </span><span style="font-size:12.8px">(10X for illumination and 20X for detection), magnification on the system is 27.7X. The illumination here is 473nm and detection at 520nm.</span></div>
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<div><span style="font-size:12.8px">A log file has been included in the zip file to give you an idea of what I've been trying. </span><br>
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<div style="font-size:12.8px">I hope I'm just missing a trick somewhere. Any help would be greatly appreciated!</div>
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<div style="font-size:12.8px">Best wishes</div>
<div style="font-size:12.8px">Tania</div>
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<div data-smartmail="gmail_signature">
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<div>Tania Mendonca
<div>Doctoral Research Student<br>
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<div>University of Sheffield<br>
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<div>S10 2TN<br>
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<div>-------------------------<br>
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</blockquote></div><br><br clear="all"><div><br></div>-- <br><div class="gmail_signature" data-smartmail="gmail_signature"><div dir="ltr">-------------------------<div>Tania Mendonca<div>Doctoral Research Student<br></div><div>University of Sheffield<br></div><div>S10 2TN<br></div></div><div>-------------------------<br></div></div></div>
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