<html><head><meta http-equiv="Content-Type" content="text/html charset=iso-8859-1"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; ">Attached is a screenshot of the issue I'm seeing, even when I deselect "compute overlap" and set the slider to 0. On the right is the LOCI stitching, which seems reasonable except the flipped y axis. The left is the output of the plugin, in which images overlap erroneously (?). When I "ignore calibration," there is no change.<div><br></div><div>Is my metadata wrong? Does the plugin handle composite (multi-channel) images well?<div><br></div><div>-Sam<br><div><br><div><div>On Jun 19, 2013, at 11:29 AM, Curtis Rueden <<a href="mailto:ctrueden@wisc.edu">ctrueden@wisc.edu</a>> wrote:</div><br class="Apple-interchange-newline"><blockquote type="cite"><div dir="ltr">Hi Sam,<div><br></div><div>> <span style="font-family:arial,sans-serif;font-size:13px">Also, what does the "ignore calibration" option do?</span></div><div><span style="font-family:arial,sans-serif;font-size:13px"><br>
</span></div><div style=""><font face="arial, sans-serif">I would test with this option both on and off. It controls whether the stage position coordinates are interpreted as pixels, or in calibrated physical units (e.g., microns).</font></div>
<div style=""><font face="arial, sans-serif"><br></font></div><div style=""><font face="arial, sans-serif">Regards,</font></div><div style=""><font face="arial, sans-serif">Curtis</font></div></div><div class="gmail_extra"><br><br>
<div class="gmail_quote">On Wed, Jun 19, 2013 at 1:27 PM, Sam Lord <span dir="ltr"><<a href="mailto:sjlord@berkeley.edu" target="_blank">sjlord@berkeley.edu</a>></span> wrote:<br><blockquote class="gmail_quote" style="margin:0 0 0 .8ex;border-left:1px #ccc solid;padding-left:1ex">
<div style="word-wrap:break-word">Thanks for the response, Curtis.<div><br></div><div>I tried setting the overlap slider to 0 and deselecting all other options. I thought this would reproduce the results that the LOCI importer stitching gave me. Instead, the final fused image was about half the dimensions, and several tiles were fused on top of each other. Maybe there's an issue reading the positions from the metadata?</div>
<div><br></div><div>I'll keep playing around with it.</div><div><br></div><div>Also, what does the "ignore calibration" option do?</div><div><br></div><div>Thanks,</div><div><br></div><div>-Sam</div><div><div class="h5">
<div><br><div><div>On Jun 19, 2013, at 9:39 AM, Curtis Rueden <<a href="mailto:ctrueden@wisc.edu" target="_blank">ctrueden@wisc.edu</a>> wrote:</div><br><blockquote type="cite"><div dir="ltr">Hi Sam,<div><br></div>
<div>
<div>> I've been having a lot of trouble getting the Fiji Grid/Collection</div><div>> Stitching plugin to operate how I want.</div><div><br></div><div>Thanks for the report. I am forwarding this thread to fiji-devel, which is a better place to report issues with Fiji plugins such as Grid/Collection Stitching.<br>
<br><div>> -If I deselect "compute overlap," I would expect the plugin to simply</div><div>> place the tiles where the stage position in the metadata claims it is.</div><div>> Instead, the output seems to not include most of the images.</div>
<div><br></div><div>Try reducing the overlap slider to 0. IIRC, it defaults to 10, and that factor still applies even if you uncheck "compute overlap."</div><div><br></div><div><div>> -When I do select "compute overlap," the images often aren't placed</div>
<div>> anywhere near the right locations. For instance, it will output an</div><div>> irregular shape, when the fused image should be a rectangle.</div><div><br></div><div>There are various ways of tuning how it approaches the computation, but if your stage coordinates are already very accurate, you probably don't need the "compute overlap" feature anyway.</div>
<div><br></div><div><div>> I'm sure some of these issues result from a non-perfect pixel</div><div>> calibration and the fact that I did not include any overlap in my</div><div>> tiles when acquiring them.</div>
<div><br></div><div>Ah, yes. If you do not include any overlap, then the "Compute overlap" feature will always fail. It relies on overlap between tiles to detect how things should be laid out. Your only option when collecting fully non-overlapping tiles is to uncheck "Compute overlap" and reduce the overlap slider to 0. It should then lay them out exactly how you collected them.</div>
</div></div><div><br></div><div>> However, the LOCI importer seems to not worry about that. Instead, it</div><div>> happy tiles my images. The only problem is that I can tell the images</div><div>> are flipped in the y direction.</div>
<div><br></div><div>Right, the LOCI importer will stitch your image *exactly* how the metadata says, and cannot do anything else such as flip tiles inverted in Y. So I suggest you try the Grid/Collection Stitching plugin again as I described above. If you still cannot get it work, and are willing to post a sample non-working dataset, we can investigate further.</div>
<div><br></div>Regards,</div><div>Curtis</div><div class="gmail_extra"><br><br><div class="gmail_quote">On Wed, Jun 19, 2013 at 11:29 AM, Sam Lord <span dir="ltr"><<a href="mailto:sjlord@berkeley.edu" target="_blank">sjlord@berkeley.edu</a>></span> wrote:<br>
<blockquote class="gmail_quote" style="margin:0px 0px 0px 0.8ex;border-left-width:1px;border-left-color:rgb(204,204,204);border-left-style:solid;padding-left:1ex"><div style="word-wrap:break-word">Hi Curtis,<div><br></div>
<div>I've been having a lot of trouble getting the Fiji Grid/Collection Stitching plugin to operate how I want. I'm asking it to select positions from file and instructing it to use the image metadata. Here are some issues I've found:</div>
<div><br></div><div>-The color scheme does not import. Neither does some other metadata (e.g. pixel size).</div><div>-If I deselect "compute overlap," I would expect the plugin to simply place the tiles where the stage position in the metadata claims it is. Instead, the output seems to not include most of the images.</div>
<div>-When I do select "compute overlap," the images often aren't placed anywhere near the right locations. For instance, it will output an irregular shape, when the fused image should be a rectangle.</div>
<div>
<br></div><div>I'm sure some of these issues result from a non-perfect pixel calibration and the fact that I did not include any overlap in my tiles when acquiring them. However, the LOCI importer seems to not worry about that. Instead, it happy tiles my images. The only problem is that I can tell the images are flipped in the y direction.</div>
<div><br></div><div>Is there a way to "brainlessly" tile my images from metadata location AND flip y?</div><span><font color="#888888"><div><br></div><div>-Sam</div></font></span><div><div>
<br></div><div><br><div><div>On Jun 16, 2013, at 11:49 AM, Curtis Rueden <<a href="mailto:ctrueden@wisc.edu" target="_blank">ctrueden@wisc.edu</a>> wrote:</div><br><blockquote type="cite"><p dir="ltr">Hi Sam,</p><p dir="ltr">
We recently added an "invert Y axis coordinates" option (and one for X as well) to the Stitching plugins. Did you try that?</p><p dir="ltr">The Fiji Grid/Collection Stitching plugin, in case you haven't tried it, uses Bio-Formats and is much more powerful than the BF Importer's basic stitching feature.</p><p dir="ltr">Regards,<br>
Curtis<br>
</p>
<div class="gmail_quote">On Jun 16, 2013 12:01 PM, "Sam Lord" <<a href="mailto:sjlord@berkeley.edu" target="_blank">sjlord@berkeley.edu</a>> wrote:<br type="attribution"><blockquote class="gmail_quote" style="margin:0px 0px 0px 0.8ex;border-left-width:1px;border-left-color:rgb(204,204,204);border-left-style:solid;padding-left:1ex">
<div><div>Importing stacks from Micro-Manager works well using Bio-Formats Importer, and the stacks option seems to read location directly from metadata. The only issue I have is that my camera must be rotated 180 degrees, because the columns stitch together well, but each row is upside-down. </div>
<div><br></div><div>I tried importing the stack, flipping, exporting, then reimorting, but something must have gotten lost in the metadata in that maneuver. </div><div><br></div><div><div>Any chance there is a simple way to tell the stitching that the images are flipped? </div>
</div><div><br>-Sam</div></div>
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