Confocal FLIM
Confocal imaging on a widefield fluorescence microscope can now be done in combination with frequency domain FLIM. The increased spatial resolution in the z-direction results in lifetime images with enhanced contrast as the detection of out-of-focus emission is reduced significantly. This allows you to see differences in fluorescence lifetime e.g. between the cell membrane and the cytoplasm. Read more about the technology of confocal imaging or view the LIFA confocal product with a photo of the set-up.
These data were obtained with the LIFA, either widefield (with LED light; 468nm peak) or confocal (with spinning disk CSU10 and 470nm-diode laser). The fluorescence lifetime images are generated at 2 different z-positions, z1 and z2. In the download more z-positions are shown, as well as avi movies through a lot of z-positions.
| Confocal (Spinning Disk) | Widefield (LED) | |
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z1 |
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z2 |
These images are extracted from the download.
| Confocal (Spinning Disk) | Widefield (LED) | |
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z1 |
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z2 |
These differences could be due to the fact that the diode laser has one excitation wavelength, exactly 470nm, while the LED has a range of wavelengths for which we used the emission band pass filter of 465-495nm.
These images are extracted from the download.
Subapplications:
Other applications in this category:
Biomolecular interactionsTIRF (Total Internal Reflection Fluorescence) - FLIM
Spectrally Resolved FLIM
Ion imaging
Probing microenvironment








