[ome-users] Two issues with ND2 (Nikon) files

David Gault (Staff) d.gault at dundee.ac.uk
Mon Sep 4 16:09:43 BST 2017


Hi Maxime,

We have been able to further debug the first issue to identify how the problem occurs but we do not yet have a fix in place for it.
You can follow the progress of that issue on the Trello card here: https://trello.com/c/RyXt2BAg/177-nd2-incorrect-channel-count

With regards the second issue I have still been unable to reproduce it. If I open the 2 converted files you had originally attached (one converted with bfconvert, the other with FIJI) both open and display the same when using the latest FIJI install.
When you converted the TIFF image using FIJI, and when you read the image back, was that using the Bio-Formats exporter and importer?

With Thanks,
David Gault

On 1 Sep 2017, at 18:59, Maxime Woringer <maxime.woringer at berkeley.edu<mailto:maxime.woringer at berkeley.edu>> wrote:

Hi David,

Do you have any update about the two bugs mentioned earlier? Were you
able to reproduce the second bug mentioned in my previous email?

Thanks a lot,
Maxime

David Gault (Staff) <d.gault at dundee.ac.uk<mailto:d.gault at dundee.ac.uk>> writes:

Hi Maxime,

Thank for reporting this issue and for providing sample files to test with.
I can certainly reproduce the first the issue using the sample files provided and this does indeed look to be a bug.
For the second issue I was unable to reproduce using bfconvert from the latest Bio-Formats release, and in theory the conversion should be the same as the FIJI result.
Would you be able to run bfconvert -version to confirm that bftools is using the latest jars?

I have created a Trello card on the Bio-Formats inbox for further investigation into these issues: https://trello.com/c/RyXt2BAg/177-nd2-incorrect-channel-count

With Thanks,
David Gault

On 14 Aug 2017, at 04:01, Maxime Woringer <maxime.woringer at berkeley.edu<mailto:maxime.woringer at berkeley.edu><mailto:maxime.woringer at berkeley.edu>> wrote:

Dear Bio-Formats users/developers,

I am trying to convert ND2 files (acquired with NIkon NIS elements) to
TIFF files, ideally in an automated fashion, but I am encountering
issues that seem to me like bugs (but I'd be happy to be proven wrong).

1. Some single images appear corrupted when I try to convert them to
TIFF. Below is an example. The two files (BF007 and BF008) were created
on the same machine, same software, at a 30' interval. I am not aware of
any differing setting except the ROI size.
- BF008.nd2 opens properly in Fiji and converts properly using bfconvert
(laters version). I am attaching the .nd2 file, the resulting TIFF file,
and the convert log (.txt files) and a screenshot of how Fiji opens it.
- BF008.nd2 opens corrupted in Fiji, and the same result happens using
bfconvert. It seems that BF thinks it is a 3-channel image (it is not).
<BF007.nd2><BF007.txt><BF007-bf.tiff><BF007-Capture du 2017-08-13 18-50-59.png><BF008.nd2><BF008.txt><BF008-bf.tiff><BF008-Capture du 2017-08-13 18-50-53.png>
2. Now moving to ND2 TIFF series (one channel, multiple timepoints),
they open fine in Fiji using BioFormats, however, if I use the following
command:

bftools/bfconvert in.nd2 out.tiff

The resulting TIFF file opens totally corrupted in Fiji/ImageJ (note
that the first frame is fine).

Below are an example source file (nd2), the conversion result using
bfconvert, and the conversion result using Fiji. I am attaching the
bfconvert log (convert.log)
<convert.log>- Source ND2 file: https://cloud.biologie.ens.fr/index.php/s/H7OPHzLvJ3MPUkd
- Corrupted TIFF generated by bfconvert: https://cloud.biologie.ens.fr/index.php/s/FFOIjHUg6Dl7B1I
- Correct TIFF generated by Fiji: https://cloud.biologie.ens.fr/index.php/s/dyl1hZha3pGiP50

Finally, I should mention that at least the latter issue happens with
files generated with several versions of NIS elements.

Am I the only person experiencing those issues? Is there any kind of
additional information I should provide?

- Version information: bf 5.5.3 and/or Fiji (latest update)
- OS: I encounter the issue on Ubuntu 14.04 and Debian Jessie (8.9)
- java version: I believe the one used is packaged by BioFormats, but in
case, java -version returns
 Ubuntu machine:
     java version "1.8.0_144"
     Java(TM) SE Runtime Environment (build 1.8.0_144-b01)
     Java HotSpot(TM) 64-Bit Server VM (build 25.144-b01, mixed mode)
 Debian machine:
     java version "1.7.0_131"
     OpenJDK Runtime Environment (IcedTea 2.6.9) (7u131-2.6.9-2~deb8u1)
     OpenJDK 64-Bit Server VM (build 24.131-b00, mixed mode)

Thanks a lot!
Maxime

PS: I tried to upload the files to the QA systemn, but it complains that
my version of Flash is not up-to-date. I apologize about that and hope
that you will be able to retrieve the files linked/attached.
PPS: I have checked the "Common issues to check" page before sending
this email, and I believe that none of them apply.
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