[ome-users] Stage positions from Zeiss LSM 710, Olympus FluoView 1000, Leica SP/SP5

Jason Swedlow jason at lifesci.dundee.ac.uk
Mon Nov 16 09:23:21 GMT 2009


Hi Aaron-

On 16 Nov 2009, at 09:09, Ponti, Aaron wrote:

> Dear Will and Jason,
>
> Thanks for your feedback. I am posting my question to the confocal   
> lists right away.
> Jason, I guess we simply don’t have a community that can make  
> pressure on vendors. I am afraid most buyers do not care much about  
> file formats and the like as long as the optics are good.

I am much more optimistic-- in the long term.  There is alot of  
interest from vendors and users-- the customers.  If we don't push  
this, the problem only gets worse.  Optics ARE important.  So is  
working with your data.

Cheers,

Jason


>
> a2
>
>  ---------------------------------------------------------------------
> | Dr. Aaron C. Ponti
> | Friedrich Miescher Institute for Biomedical Research
> | Facility for Advanced Microscopy and Imaging
> | Software development
> | Maulbeerstrasse 66 CH-4058, Basel
> | WRO-1066.2.16
> | Tel: +41 61 696 3513
> | Fax: +41 61 697 3976
> | http://www.fmi.ch/faim
>   
> ----------------------------------------------------------------------
>
> From: Jason Swedlow [mailto:jason at lifesci.dundee.ac.uk]
> Sent: Saturday, November 14, 2009 9:40 AM
> To: Ponti, Aaron
> Cc: ome-users at lists.openmicroscopy.org.uk OME-Users
> Subject: Re: [ome-users] Stage positions from Zeiss LSM 710, Olympus  
> FluoView 1000, Leica SP/SP5
>
> Hi Aaron-
>
> Just adding one comment.  From our contacts with Carl Zeiss  
> MicroImaging we've been informally told that there were no changes  
> in the LSM 710's file formats that we needed to know about.   
> However, these formats are very complex, and something might have  
> slipped by.
>
> As Will mentioned, the confocal list server might be a good place to  
> post this (this was the method we used some time ago).
>
> OK, climbing on soapbox:
>
> The current situation-- arbitrary file formats, with essentially any  
> metadata in any format, and no definitive community notification  
> when things change-- will only change when customers condition their  
> purchases on this information being provided.  In an ideal world  
> (ok, a total pipe dream), our OME-XML and Bio-Formats developers  
> would be notified BEFORE new formats were released and provided  
> specifications and samples.  We're a long way from that, but can get  
> there, with the community's help.  If this community insists that  
> this information will be provided, we won't be having these problems  
> anymore.
>
> Stepping down.  Phew!
>
> Have a great weekend.
>
> Cheers,
>
> Jason
>
>
> On 13 Nov 2009, at 17:28, Will Moore wrote:
>
>
> Hi Aaron,
>
> The key question is whether the stage position metadata is actually  
> stored in the file. If it is, then it should be possible for  
> BioFormats to identify the correct bit of metadata and store it in  
> the OME data model.
>
> We have been working at improving the amount of metadata from these  
> confocal formats that is used to populate the OME model. This takes  
> time to identify how to read the piece of metadata in the original  
> file, and how to map it into the OME model. However, we have so far  
> been successful for the various bits of metadata we've attempted so  
> far.
>
> There are a couple of ways to determine whether the stage position  
> metadata is actually in the file:
> Open the file in the proprietary software (Olympus, Zeiss, Lecia)  
> and see if it is displayed.
> Or, open the file in ImageJ, using the BioFormats plugin and look at  
> the "Original Metadata".
>
> I've tried these with a sample of the various formats you mentioned,  
> and I don't see that any of them have absolute stage positions.  
> (some examples below)
> I'm not even sure that BioFormats is reading the Leica stage  
> positions properly. Haven't looked at this before.
>
> However, I don't have any LSM 710 files (LSM 510 only), and it's  
> also possible that newer versions of these microscopes might save  
> this data.
> Some of these files seem to have places in the files for this  
> metadata, and it's also possible that the formats might expand to  
> accommodate stage metadata.
>
> You could try asking on the confocal lists http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>
>   Sorry I couldn't be more help,
>
>   Will.
>
>
>
> Looking at the "original metadata" of a couple of oib files, read by  
> Bioformats (using the ImageJ plugin). NB. This is all the  
> "available" metadata currently read by BioFormats, only a subset of  
> this is used to populate the OME data model.
> This is the data available for each plane of the image, shown for  
> the first 2 planes of a Z stack.
>
> Image 0 : AS Level       9999
> Image 0 : AbsPositionUnitName           mm
> Image 0 : AbsPositionValue      0.0
> Image 0 : Adjust Outward         0
> Image 0 : Adjust Retrun            0
> Image 0 : AnalogPMTGain       1.0
> Image 0 : AnalogPMTOffset     2
> Image 0 : Confocal       ON
> Image 0 : CountingPMTGain     0.0
> Image 0 : CountingPMTOffset  1.600000
> Image 0 : CountingPMTVoltage            0
> Image 0 : DataMax       4095.0
> Image 0 : DataMin        0.0
> Image 0 : DataName     1h_after_pdt_C001Z001T001.tif
> Image 0 : DataType      WORD
> Image 0 : ExcitationOutPutLevel           5
> Image 0 : HeightConvertValue  0.207
> Image 0 : HeightUnit     um
> Image 0 : ImageDepth   2
> Image 0 : ImageGroup  Normal
> Image 0 : ImageHeight  1024
> Image 0 : ImageType    Intensity
> Image 0 : ImageWidth   1024
> Image 0 : LUTFileName           LUT1
> Image 0 : LUTParameter          530227280
> Image 0 : LightControl  9999
> Image 0 : Magnification 60.0
> Image 0 : Name            COFAFrameImage
> Image 0 : Number         1
> Image 0 : ObjectiveLens NAValue        1.35
> Image 0 : ObjectiveLens Name UPLSAPO  60X O  NA:1.35
> Image 0 : ObjectiveLens WDValue       9999.0
> Image 0 : Observation Mode    LSM
> Image 0 : PMTDetectingMode  Analog
> Image 0 : PMTVoltage 830
> Image 0 : Pan Scale      2
> Image 0 : Path  .\
> Image 0 : PinholeDiameter        115000
> Image 0 : PinholeScale  1
> Image 0 : PixConvertValue       1.0
> Image 0 : PixUnit          Intensity
> Image 0 : ROIFileName            530223744_9038484
> Image 0 : Resolution     10.0
> Image 0 : RotationValue            1.0
> Image 0 : RotationValue After Clip        0
> Image 0 : RotationValue Before Clip     10
> Image 0 : SamplingClock          500000
> Image 0 : ScanSpeed    2.0
> Image 0 : Step  0.0
> Image 0 : ValidBitCounts          12
> Image 0 : Version         1.0.0.0
> Image 0 : WidthConvertValue   0.207
> Image 0 : WidthUnit      um
> Image 0 : X Pinhole      318
> Image 0 : XPanValue    0
> Image 0 : Y Pinhole      -570
> Image 0 : YPanValue    0
> Image 0 : ZoomValue   1.0
>
> Image 1 : AS Level       9999
> Image 1 : AbsPositionUnitName           mm
> Image 1 : AbsPositionValue      0.0
> Image 1 : Adjust Outward         0
> Image 1 : Adjust Retrun            0
> Image 1 : AnalogPMTGain       1.0
> Image 1 : AnalogPMTOffset     3
> Image 1 : Confocal       ON
> Image 1 : CountingPMTGain     0.0
> Image 1 : CountingPMTOffset  1.600000
> Image 1 : CountingPMTVoltage            0
> Image 1 : DataMax       4095.0
> Image 1 : DataMin        0.0
> Image 1 : DataName     1h_after_pdt_C002Z001T001.tif
> Image 1 : DataType      WORD
> Image 1 : ExcitationOutPutLevel           4
> Image 1 : HeightConvertValue  0.207
> Image 1 : HeightUnit     um
> Image 1 : ImageDepth   2
> Image 1 : ImageGroup  Normal
> Image 1 : ImageHeight  1024
> Image 1 : ImageType    Intensity
> Image 1 : ImageWidth   1024
> Image 1 : LUTFileName           LUT2
> Image 1 : LUTParameter          530238208
> Image 1 : LightControl  9999
> Image 1 : Magnification 60.0
> Image 1 : Name            COFAFrameImage
> Image 1 : Number         1
> Image 1 : ObjectiveLens NAValue        1.35
> Image 1 : ObjectiveLens Name UPLSAPO  60X O  NA:1.35
> Image 1 : ObjectiveLens WDValue       9999.0
> Image 1 : Observation Mode    LSM
> Image 1 : PMTDetectingMode  Analog
> Image 1 : PMTVoltage 700
> Image 1 : Pan Scale      2
> Image 1 : Path  .\
> Image 1 : PinholeDiameter        115000
> Image 1 : PinholeScale  1
> Image 1 : PixConvertValue       1.0
> Image 1 : PixUnit          Intensity
> Image 1 : ROIFileName            530223744_9038484
> Image 1 : Resolution     10.0
> Image 1 : RotationValue            1.0
> Image 1 : RotationValue After Clip        0
> Image 1 : RotationValue Before Clip     10
> Image 1 : SamplingClock          500000
> Image 1 : ScanSpeed    2.0
> Image 1 : Step  0.0
> Image 1 : ValidBitCounts          12
> Image 1 : Version         1.0.0.0
> Image 1 : WidthConvertValue   0.207
> Image 1 : WidthUnit      um
> Image 1 : X Pinhole      318
> Image 1 : XPanValue    0
> Image 1 : Y Pinhole      -570
> Image 1 : YPanValue    0
> Image 1 : ZoomValue   1.0
>
> It looks like there might be some parameters for storing stage  
> position (e.g. YPanValue) but they are not populated.
> I also don't see any stage position info displayed in the Olympus  
> software for this file, so I'm assuming that it is not stored.
> However, I don't have a file format specification to hand, and it's  
> also possible that newer microscopes might store this data.
>
>
> Looking at a couple of Leica LIF files, I see that the metadata  
> displayed by the software does not include stage position, but if I  
> look in the exported metadata, I can see StagePosX="0" StagePosY="0"  
> StagePosZ="0". So, the parameters exist in the file but it is not  
> written in this case. This may be because the microscope that  
> acquired this image was not fitted with a motorized stage?
>
>
>
> On 13 Nov 2009, at 16:05, Ponti, Aaron wrote:
>
>
> Hello
>
> We are investigating the purchase of one of the following confocals:  
> Zeiss LSM 710, Olympus FluoView 1000, and Leica SP/SP5. One of the  
> criteria for the choice is the possibility to read stage positions  
> from the files (using the loci/ome-xml tools). With loci_tools 4.1  
> it seems that reading the stage positions into the OME schema is  
> supported for the Leica, but is not for the other two. Can anybody  
> confirm this? Does anybody know if stage positions arestored at all  
> in .lsm and .oib files?
>
> Thanks
>
>  ---------------------------------------------------------------------
> | Dr. Aaron C. Ponti
> | Friedrich Miescher Institute for Biomedical Research
> | Facility for Advanced Microscopy and Imaging
> | Software development
> | Maulbeerstrasse 66 CH-4058, Basel
> | WRO-1066.2.16
> | Tel: +41 61 696 3513
> | Fax: +41 61 697 3976
> | http://www.fmi.ch/faim
>   
> ----------------------------------------------------------------------
>
>
> _______________________________________________
> ome-users mailing list
> ome-users at lists.openmicroscopy.org.uk
> http://lists.openmicroscopy.org.uk/mailman/listinfo/ome-users
>
> William Moore
> Division of Gene Regulation and Expression
> College of Life Sciences
> University of Dundee
> Scotland
> DD1 5PH
>
> Tel 01382 386364
>
> _______________________________________________
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> ome-users at lists.openmicroscopy.org.uk
> http://lists.openmicroscopy.org.uk/mailman/listinfo/ome-users
>
>
>
> **************************
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> College of Life Sciences
> MSI/WTB/JBC Complex
> University of Dundee
> Dow Street
> Dundee  DD1 5EH
> United Kingdom
>
> phone (01382) 385819
> Intl phone:  44 1382 385819
> FAX   (01382) 388072
> email: jason at lifesci.dundee.ac.uk
>
> Lab Page: http://gre.lifesci.dundee.ac.uk/staff/jason_swedlow.html
> Open Microscopy Environment: http://openmicroscopy.org
> **************************
>
> The University of Dundee is a Scottish Registered Charity, No.  
> SC015096.
>
>
>



**************************
Wellcome Trust Centre for Gene Regulation & Expression
College of Life Sciences
MSI/WTB/JBC Complex
University of Dundee
Dow Street
Dundee  DD1 5EH
United Kingdom

phone (01382) 385819
Intl phone:  44 1382 385819
FAX   (01382) 388072
email: jason at lifesci.dundee.ac.uk

Lab Page: http://gre.lifesci.dundee.ac.uk/staff/jason_swedlow.html
Open Microscopy Environment: http://openmicroscopy.org
**************************

The University of Dundee is a Scottish Registered Charity, No. SC015096.



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