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Fluorescence Lifetime Imaging

Fluorescence lifetime imaging microscopy (FLIM) is an imaging technique that makes use of the inherent properties of fluorescent dyes. Aside from having characteristic emission spectra, each fluorescent molecule has a characteristic lifetime that reflects how long the fluorophore spends in the excited state before emitting a photon. Lifetime analysis provides you with information in addition to your standard fluorescence intensity measurements.

Historically, lifetime imaging has been a slow, complex, and specialized technique. Its use was limited to experienced microscopists or physicists. Leica Microsystems is at the cutting edge of today’s fluorescence lifetime imaging advances. Our systems make lifetime imaging faster and easier to use than ever before, binging its advantages to every day confocal imaging experiments.

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Boost confocal imaging with fluorescence lifetime information

Fluorescence lifetime imaging provides additional information that can help you improve the quality of your confocal imaging. It can be particularly useful to discriminate fluorescence probes that have overlapping fluorescent emission spectra, or to eliminate unwanted background fluorescent signals.

FLIM is a crucial application for measuring and quantifying imaging data. As lifetime information is independent of fluorophore concentration, it is extremely well suited for functional imaging. Functional imaging goes beyond the traditional recording of the location and concentration of molecular species and enables further investigation of molecular function, interactions, and their environment. 

Additionally, FLIM enables novel imaging applications, such as the use of biosensors to study the cell’s microenvironment or imaging based on the intrinsic fluorescent properties of your sample.

Learn more about FLIM and its imaging applications with the SP8 FALCON system by referring to this application note.

The lifetime contrast of sepal of silverberry allows to separate different structure within the sample. The intensity image (yellow image) doesn't show any specific structure while the FastFLIM (rainbow scale image) and the lifetime fitted images (RGB, three component image) give insigth into the different part of the sample. The end of the video shows the three structures determined by the lifetime fit split into three channels (RGB).

Setting the standard for lifetime imaging

With the release of the fast and fully integrated lifetime imaging technology – the SP8 FALCON – Leica Microsystems set the benchmark for the future of lifetime imaging. Leica Microsystems received the prestigious R&D 100 Award for the SP8 FALCON system. This award recognizes the solution’s innovative approach to lifetime imaging.

Leica Microsystems solutions for FLIM enable you to harness the power of fluorescence lifetime to extract a new dimension of information from every sample with your confocal experiment. Investigate cellular physiology and explore cellular dynamics with FALCON. Get immediate access to functional information such as metabolic status, pH and ion concentration with the potential of Tausense in STELLARIS.

With our simplified, integrated platforms, you can gain new perspectives on your experiments, allowing you to harness new answers to your experimental questions.

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Mouse embryo mosaic image

High resolution mouse embryo mosaic image of 722 tiles containing 190 Megapixels. FLIM data fitted with four characteristic fluorescence lifetimes, color coded. Acquisition: 1:23 h. Analysis: 1:00 h

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