NovaFLIM can export FLIM images as stacked TIFFs containing multiple image layers (e.g., Intensity + Lifetime). This article explains how to correctly open and interpret these files in FIJI/ImageJ.

Opening the TIFF in FIJI/ImageJ

Opening the stacked TIFF requires the Bio-Formats Importer plugin in FIJI.

1. Open the TIFF in FIJI using: Plugins → Bio-Formats → Bio-Formats Importer. In the import dialog, enable Display metadata and Display OME-XML metadata.

2. In the OME Metadata window, check the Channel@ID entries. For example, Channel:0:0... refers to the first layer (leftmost in the TIFF viewer), which typically corresponds to the "Intensity (FLIM)" image. Subsequent layers represent additional channels such as "Fast Lifetime (FLIM)", "Slow Lifetime", etc.

Troubleshooting

If the Bio-Formats Importer plugin does not work as expected, this may be due to a version incompatibility. Please make sure you are using the latest version of FIJI/ImageJ, which can be downloaded from https://imagej.net/downloads.

First reported: to be added

Linked Tickets: to be added