[ome-users] Fiji Bio-Formats Exporter Problem - write each timepoint to a separate file

Wed Dec 14 08:10:19 GMT 2016

Hi,

Thanks for looking into this.
I tried to reproduce the problem and it is different from what I wrote before.

In Summary:
1. separating into different ome.tif files for each time points - works OK
2. separating into different tif files for each time points - produce file with corrupted pixel values (only first slice is OK)
3. saving hyperstack into one ome.tif produce file which is missing one image and cannot be rearranged into hyperstack correctly.
4. saving hyperstack into one tif produce file of the correct size but with corrupted pixel values (only first slice is OK)
5. saving hyperstack into one tif with regular Fiji Save As tif works fine

I uploaded sample files for you into my box folder: https://weizmann.box.com/s/xhbixjp0ydbd5qd69s2xnm9k2lugvdri
It also reproduced with the Mitosis sample file

See details below

Best Regards
Ofra

Details:
=====

BioFormats version: 5.3.0-SNAPSHOT,  Build Date: 9 decemebr 2016

Here is my recorded command when saving in tif format
        run("Bio-Formats Exporter", "save=D:\\UserData\\innaa\\OfraTest\\dlGFP_Test_NewFiji\\Mitosis\\mito1.tif write_each_timepoint compression=Uncompressed");

I opened the file of the first time-point ("mito1_T0.tif")
Attached are snapshots comparing the slices to the original slices and the corrupted file itself

Here is the recorded command for saving in ome.tif format
        run("Bio-Formats Exporter", "save=D:\\UserData\\innaa\\OfraTest\\dlGFP_Test_NewFiji\\Mitosis\\mito2.ome.tif write_each_timepoint compression=Uncompressed");

When I try to open it by drag and drop the file ("mito2_T0.ome.tif")  is opened as stack instead of hyperstack , after reorganizing it, it looks OK (Can't reproduce the corruption).
When I try to open it with BioFormats Importer I get an exception (attached)

The same behavior reproduced on my workstation (Windows7, which has Release 5.3.0-SNAPSHOT build date 5 December 2016, see attachment)

My original problem is with much bigger czi file (3D+time) that I try to crop and output to different files for different time-points.

I crop part of the big file (95 GB) so that I have now 16 slices * 11 time points

- exporting (on the WindowsServer WS) to ome.tif produces a file with only 175 images so 1 image is missing and I cannot rearrange it into hyperstack:
run("Bio-Formats Exporter", "save=D:\\UserData\\innaa\\OfraTest\\dlGFP_Test_NewFiji\\tmp\\AllTP.ome.tif compression=Uncompressed");

- exporting into into one tif file produces hyperstack of the correct size, but with corrupted pixel values

run("Bio-Formats Exporter", "save=D:\\UserData\\innaa\\OfraTest\\dlGFP_Test_NewFiji\\tmp\\AllTP.tif compression=Uncompressed");

- Saving the file using the regular Fiji Save As Tif option produces a hyperstack of the real size with the original pixel values - You can use it as a starting point for reproducing the problems

saveAs("Tiff", "D:\\UserData\\innaa\\OfraTest\\dlGFP_Test_NewFiji\\tmp\\AllTP_SaveAsTif.tif");

However - I need to separate into independent file for each time point. So when using BioFormat Exporter to save each independent TP into file with

run("Bio-Formats Exporter", "save=D:\\UserData\\innaa\\OfraTest\\dlGFP_Test_NewFiji\\tmp\\SepTP.ome.tif write_each_timepoint compression=Uncompressed");

produce valid stacks - so no problem here

When writing into tif the output files have corrupted pixel values

run("Bio-Formats Exporter", "save=D:\\UserData\\innaa\\OfraTest\\dlGFP_Test_NewFiji\\tmp\\SepTP.tif write_each_timepoint compression=Uncompressed");

-----Original Message-----
From: ome-users [mailto:ome-users-bounces at lists.openmicroscopy.org.uk] On Behalf Of Balaji Ramalingam (Staff)
Sent: Tuesday, December 13, 2016 5:56 PM
To: OME User Support List
Subject: Re: [ome-users] Fiji Bio-Formats Exporter Problem - write each timepoint to a separate file

Hi Ofra,

Thank you for reporting your issue.
Testing the same locally, we were unfortunately unable to reproduce the bug.

I have recorded the Fiji/ImageJ operations that I had performed (using the Fiji/ImageJ macro recorder) and I have pasted the same here,

run("Bio-Formats Exporter",
"save=/Users/bramalingam/Desktop/FijiExportTest/mitosisOmeTiff/mitosis_time
point_.ome.tif write_each_timepoint compression=Uncompressed²);

I reimported the resultant images back into Fiji/ImageJ and compared the pixel data with the original image.

There was no obvious issue that was noticed, thus could you please record your mouse-click operations via the Fiji/ImageJ macro recorder and paste the resultant command here? The documentation for the same can be found here, https://imagej.net/Introduction_into_Macro_Programming#The_recorder

And also it would be great if you could clarify the version of Bio-Formats that was used? (I have updated my Fiji to use the latest release version of Bio-Formats : Version 5.3.0)

You can get the same by selecting,
Help‹>About Plugins‹>Bio-Formats PluginsŠ (in the Fiji menu)

Or by running the following macro command, run("Bio-Formats Plugins...²);

This would help us troubleshoot the issue in better detail.

Best,
Balaji

__________________
Mr Balaji Ramalingam
Software Developer

OME Team
School of Life Sciences
University of Dundee

On 12/12/2016, 14:54, "ome-users on behalf of Ofra Golani"
<ome-users-bounces at lists.openmicroscopy.org.uk on behalf of ofra.golani at weizmann.ac.il> wrote:

>Hi,
>
>I encounter a problem when using Bio-Formats exporter to save each
>time-point into separate file.
>It saves into separate files, but the files are corrupted. You see this
>when you move through the slices.
>You can try this with the "Mitosis" 5D stack of Fiji Sample files.
>The problem happens when I save files both as tif and as ome.tif.
>I use the latest Fiji version, with the latest Bio-Formats version.
>
>Please advise,
>Thanks,
>
>Ofra
>
>----------------------------------------
>
>Ofra Golani
>
>BioImage Analyst
>Life Sciences Core Facilities
>Weizmann Institute of Science
>Tel: +972-8-934-5177
>ofra.golani at weizmann.ac.il
>http://www.weizmann.ac.il/Biological_Services/units/cell-observatory/ab
>out
>-service
>
>
>_______________________________________________
>ome-users mailing list
>ome-users at lists.openmicroscopy.org.uk
>http://lists.openmicroscopy.org.uk/mailman/listinfo/ome-users

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