What is FLIM?
Fluorescence Lifetime Imaging Microscopy or FLIM is an imaging technique for producing an image based on the differences in the exponential decay rate of the fluorescence from a fluorescent sample. It can be used as an imaging technique in confocal microscopy, Two-photon excitation microscopy, and multiphoton tomography.
The lifetime of the fluorophore signal, rather than its intensity, is used to create the image in FLIM. This has the advantage of minimizing the effect of photon scattering in thick layers of sample
Typical FLIM applications
Non-stained samples (autofluorescence):
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Metabolic state characterization
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Cancer research: identification of modified cells
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Plant physiology: pathogen-host interaction
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Morphology: Identification of chitin or lignin structures
Stained samples:
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Energy transfer (FRET) for molecule binding and distance measurements
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Local concentration of ions (Ca2+, Na+, pH, ...), small ligands, oxygen
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Environmental studies (viscosity, refractive index, membrane potential)
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Dye separation
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Photophysical dye properties
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"The SuperK provides a flexible laser source that allows selecting the excitation band as a tunable laser. With the SuperK you can tune in to the excitation optimum where the conventional lasers lines are unable to reach."
Dr. Bernd Sagmuller, Head of Business Segment Confocal Imaging, Leica Microsystems, Mannheim Germany
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What do I need?
Building an fluorescent imaging setup with a SuperK source is simple. All you need is a SuperK laser source, a fiber delivery set and one of our plug&play computer controlled filters:
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If you are unsure of which SuperK laser to get or which of the filters you need, don't worry. Our sales team is very experienced and has helped many customers needing a solution for fluorescence imaging.
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FLIM image of scale insect antenna (non-stained). Image was reconstructed from a 3-dimensional FLIM-z-stack. Image courtesy of Kees Jalink
How can I use a SuperK supercontinuum laser for FLIM?
Until now, FLIM setups have been limited by the type or number of dyes that could be excited. Conventional Fluorescence Lifetime Imaging Microscopes are typically equipped with a set of gas or solid state lasers that emit wavelength-fixed laser lines. Using this kind of equipment one has to make compromises by choosing the dyes and the labels according to the available wavelengths and more often than not, the available wavelengths does not match the absorption peak of the fluorescent dyes.
The SuperK supercontinuum laser eliminates all the usual compromises. The SuperK emits a broad spectrum ranging from blue and into the near infrared and in combination with a tunable filter, the excitation source can now be matched perfectly to the absorption of any dye. This has several advantages:
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Only one source needed to cover all available dyes
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No alignment of multiple sources leading to better stability and less maintenance
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Excitation can be tuned to the absorption peak of the dye maximizing the signal-to-noise ratio.
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Less power is needed as excitation source can be tuned to the absorption peak
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Less cross excitation in samples with several dyes
Moreover, the SuperK EXTREME comes with a pulse picker option that enables the repetition rate to be adjusted on the fly so that you can adjust the excitation to the decay time of your sample while the system is running.
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