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.
Our experts on solutions for FLIM are happy to help you with their advice.
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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Related Articles
Read our latest articles about Fluorescence Lifetime Imaging
The knowledge portal of Leica Microsystems offers scientific research and teaching material on the subjects of microscopy. The content is designed to support beginners, experienced practitioners and scientists alike in their everyday work and experiments.
Fluorescence Lifetime-based Imaging Gallery
Phasor FLIM Detects Metabolic Differences
Accessing the Metabolic Information of Stem Cells
How to Quantify Changes in the Metabolic Status of Single Cells
Label-free Differentiation of Microplastics
Adding Dimensions to Multiplex Molecular Imaging
How FLIM Microscopy helps to detect microplastic pollution
Advanced Quantitative Fluorescence Microscopy to Probe the Molecular Dynamics of Viral Entry
Time-resolved STED microscopy
Imaging Intracellular Temperature using Fluorescence Lifetime Imaging Microscopy (FLIM)
SP8 FALCON: a novel concept in fluorescence lifetime imaging enabling video-rate confocal FLIM
Development of fluorescence lifetime imaging microscopy (FLIM) and its relevance for functional imaging
How FRET/FLIM Biosensors and Lifetime Imaging Helped Identify the Importance of Zinc for Human B-Cell Development
Single-residue bioorthogonal labeling of G protein-coupled receptors (GPCRs) in live cells for quantitative analysis
Phasor Analysis for FLIM (Fluorescence Lifetime Imaging Microscopy)
Researchers use Functional Imaging to Elucidate the Role of Cellular Zn²⁺ in Human Immunity
SP8 DIVE Multiphoton Microscope Image Gallery
Precise and lightning-fast spectral Fluorescence Lifetime Imaging at video rate integrated in a high-end confocal microscope
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