ImageJ is an image processing and analysis application written in Java, widely used in the life sciences fields, with an extensible plugin infrastructure. You can use Bio-Formats as a plugin for ImageJ to read and write images in the formats it supports.
Download bioformats_package.jar and drop it into your ImageJ/plugins folder. Next time you run ImageJ, a new Bio-Formats submenu with several plugins will appear in the Plugins menu, including the Bio-Formats Importer and Bio-Formats Exporter.
The Bio-Formats Importer plugin can display image stacks in several ways:
In a standard ImageJ window (including as a hyperstack)
Using the LOCI Data Browser plugin (included)
With Joachim Walter’s Image5D plugin (if installed)
With Rainer Heintzmann’s View5D plugin (if installed)
ImageJ v1.37 and later automatically (via
the Bio-Formats logic, if installed, as needed when a file is opened
within ImageJ, i.e. when using instead of
For a more detailed description of each plugin, see the Bio-Formats page of the ImageJ wiki.
To upgrade, just overwrite the old bioformats_package.jar with the latest one.
You can also upgrade the Bio-Formats plugin directly from ImageJ. Selectfrom the ImageJ menu, then select which release you would like to use. You will then need to restart ImageJ to complete the upgrade process.
Macros and plugins¶
Bio-Formats is fully scriptable in a macro, and callable from a plugin. To use in a macro, use the Macro Recorder to record a call to the Bio-Formats Importer with the desired options. You can also perform more targeted metadata queries using the Bio-Formats macro extensions.
Here are some example ImageJ macros and plugins that use Bio-Formats to get you started:
basicMetadata.txt - A macro that uses the Bio-Formats macro extensions to print the chosen file’s basic dimensional parameters to the Log.
planeTimings.txt - A macro that uses the Bio-Formats macro extensions to print the chosen file’s plane timings to the Log.
recursiveTiffConvert.txt - A macro for recursively converting files to TIFF using Bio-Formats.
bfOpenAsHyperstack.txt - This macro from Wayne Rasband opens a file as a hyperstack using only the Bio-Formats macro extensions (without calling the Bio-Formats Importer plugin).
zvi2HyperStack.txt - This macro from Sebastien Huart reads in a ZVI file using Bio-Formats, synthesizes the LUT using emission wavelength metadata, and displays the result as a hyperstack.
dvSplitTimePoints.txt - This macro from Sebastien Huart splits timepoints/channels on all DV files in a folder.
batchTiffConvert.txt - This macro converts all files in a directory to TIFF using the Bio-Formats macro extensions.
Read_Image - A simple plugin that demonstrates how to use Bio-Formats to read files into ImageJ.
Mass_Importer - A simple plugin that demonstrates how to open all image files in a directory using Bio-Formats, grouping files with similar names to avoiding opening the same dataset more than once.
“How do I make the options window go away?” is a common question. There are a few ways to do this:
To disable the options window only for files in a specific format, select, then pick the format from the list and make sure the “Windowless” option is checked.
To avoid the options window entirely, use themenu item to import files.
Open files by calling the Bio-Formats importer plugin from a macro.
A common cause of problems having multiple copies of
bioformats_package.jarin you ImageJ plugins folder, or a copy of
bioformats_package.jarand a copy of
formats-gpl.jar. It is often difficult to determine for sure that this is the problem - the only error message that pretty much guarantees it is a
NoSuchMethodException. If you downloaded the latest version and whatever error message or odd behavior you are seeing has been reported as fixed, it is worth removing all copies of
loci_tools.jaror any other Bio-Formats jars) and download a fresh version.
The Bio-Formats Exporter plugin’s file chooser will automatically add the first listed file extension to the file name if a specific file format is selected in the
Files of Typebox (e.g.
.ome.tiffor OME-TIFF). This can prevent BigTIFF and OME BigTIFF files from being created, as the
.ome.btffile extension will be overwritten. To ensure that the desired extension is used, select or in the
Files of typebox, as an extension will not be automatically added in those cases.
Saving an open image using Bio-Formats must be done via TIFF (Tagged Image File Format) for additional information.or the corresponding macro code. and do not use Bio-Formats. In particular, using to save a TIFF will result in an ImageJ-specific TIFF being written. While Bio-Formats can read ImageJ TIFFs, other software may not; see