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How to Quantify Changes in the Metabolic Status of Single Cells
Metabolic imaging based on fluorescence lifetime provides insights into the metabolic dynamics of cells, but its use has been limited as expertise in advanced microscopy techniques was needed.
Now,…
How FLIM Microscopy Helps to Detect Microplastic Pollution
The use of autofluorescence in biological samples is a widely used method to gain detailed knowledge about systems or organisms. This property is not only found in biological systems, but also…
Step by Step Guide for FRAP Experiments
Fluorescence Recovery After Photobleaching (FRAP) has been considered the most widely applied method for observing translational diffusion processes of macromolecules. The resulting information can be…
Widefield Calcium Imaging with Calcium Indicator Fura2
In eukaryotic cells Ca2+ is one of the most widespread second messengers used in signal transduction pathways. Intracellular levels of Ca2+ are usually kept low, as Ca2+ often forms insoluble…
Fluorescence Correlation Spectroscopy (FCS)
Fluorescence correlation spectroscopy (FCS) measures fluctuations of fluorescence intensity in a sub-femtolitre volume to detect such parameters as the diffusion time, number of molecules or dark…
Fluorescence Recovery after Photobleaching (FRAP) and its Offspring
FRAP (Fluorescence recovery after photobleaching) can be used to study cellular protein dynamics: For visualization the protein of interest is fused to a fluorescent protein or a fluorescent dye. A…
Förster Resonance Energy Transfer (FRET)
The Förster Resonance Energy Transfer (FRET) phenomenon offers techniques that allow studies of interactions in dimensions below the optical resolution limit. FRET describes the transfer of the energy…