[ome-users] Support for .FLI files from Capture software (HiCAM FLUO high-speed camera)

[Sebastien Besson (Staff)]

Hi Aaron,

On 14 Dec 2018, at 10:00, Ponti Aaron <aaron.ponti at bsse.ethz.ch<mailto:aaron.ponti at bsse.ethz.ch>> wrote:

Hello,

we have .FLI files from Lambert Instruments' Capture software (version 1.0.4.0) for the high-speed camera HiCAM FLUO. Images are streamed at high speed and stored as UINT12 (2 pixels in 3 bytes) or optionally as UINT8. The FLIM reader in bio-formats (https://github.com/openmicroscopy/bioformats/blob/develop/components/formats-gpl/src/loci/formats/in/LiFlimReader.java) does not seem to support 12-bit intensities.

From the internal specification of the LI-FLIM file format which we used for the reader [1]:

"The datatype key can have one of the following 8 values: UINT8, INT8, UINT16, INT16,
UINT32, INT32, REAL32 or REAL64.”

A file that would use UINT12 would probably be a new variant of the file format for which we
have currently neither a format specification nor representative samples

Also, when asking bio-formats to return the metadata, the whole file is returned (which suggests that the metadata of standard FLIM .FLI files is organized -- or terminated -- differently).

Understood. This would be consistent with the pixel type issue reported above i.e. that could
indeed reflect an internal layout different from the one Bio-Formats is expecting..

 The file is binary with the first bytes containing metadata information in readable text format. One field is 'hasDarkImage'. If hasDarkImage == 1, the very last frame is a dark image that is subtracted on-the-fly in the software when displaying the frames (but the images are stored raw in the file). The 'exposureTime' is in micro seconds. Finally, the software seems to support 'compression', although there is no setting for turning it on in the software. The software supports binning.

 Would you be interested in adding support for these files to bio-formats? We have example files that we could upload.

Over the past years, we have been working on ways to have more sustainable support for new file
formats including any variants of existing file formats. We published a blog post summarizing our
experience a few years ago [2] and since then more manufacturers have been engaging in this
process.

Have you been in touch with Lambert Instruments to report this issue? Are they aware that the latest files
produced by their software are no longer readable by Bio-Formats and any software based upon it?

Representative samples allowing to reproduce the issue are always welcome. Ideally public samples licensed
under CC-BY license [3] are the most effective way to help spontaneous or sponsored contributions to the
project and facilitate both the immediate software work as well as the long-term maintenance.
You can upload datasets under 2GB to our QA system [4] and we can provide FTP credentials for large samples.

Best,
Sebastien


[1] https://docs.openmicroscopy.org/bio-formats/5.9.2/formats/lambert-instruments-flim.html
[2] https://blog.openmicroscopy.org/file-formats/community/2016/08/31/bf-partnerships/
[3] https://downloads.openmicroscopy.org/images/
[4] http://qa.openmicroscopy.org.uk/qa/upload/


 Thanks a lot,
a2


----
Dr. Aaron Ponti
Software and Data Management Engineer
Image Analysis Specialist
Single Cell Facility
Department of Biosystems Science and Engineering (D-BSSE)
ETH Zürich
Office 2.30
Mattenstrasse 26
4058 Basel, Switzerland
aaron.ponti at bsse.ethz.ch<mailto:aaron.ponti at bsse.ethz.ch>
T: +41 61 387 33 74
F: +41 61 387 39 93

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