Phosphorescence Lifetime Imaging
Phosphorescence Lifetime Imaging - Related Videos
10:38
Synthesis and Calibration of Phosphorescent Nanoprobes for Oxygen Imaging in Biological Systems
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Cited by 2 •
2010
We present principles of oxygen measurements by phosphorescence quenching and review design of porphyrin-based dendritic nanosensors for oxygen imaging in biological systems.
09:45
Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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Cited by 17 •
2012
Fluorescence Lifetime Imaging (FLIM) has emerged as a key technique to image the environment and interaction of specific proteins and dyes in living cells. FLIM of fluorescent molecular rotors allows mapping of viscosity in living cells.
03:26
Analyzing Amyloid Structures in a Tissue Section Using Fluorescence Lifetime Imaging Microscopy
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2025
Source: Nyström, S., et al., Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging. J. Vis. Exp. (2017).This video demonstrates analyzing amyloid structures with fluorescence lifetime imaging microscopy or FLIM, using hFTAA dye to reveal compact cores as blue/green and unstable peripheries as red/yellow in color-coded images, reflecting amyloid organization.
08:58
Cerebral Blood Oxygenation Measurement Based on Oxygen-dependent Quenching of Phosphorescence
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Cited by 28 •
2011
We present an experimental procedure for measuring the partial pressure of oxygen (pO2) in cerebral vasculature based on oxygen-dependent quenching of phosphorescence. Animal preparation and imaging procedures were outlined for both large field of view CCD-based imaging of pO2 in rats and 2-photon excitation based imaging of pO2 in mice.
05:36
Fluorescence Lifetime Imaging of PolyQ Protein Aggregation in Caenorhabditis elegans Neurons
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2025
Source: Pigazzini, M. L., et. al. Characterization of Amyloid Structures in Aging C. Elegans Using Fluorescence Lifetime Imaging. J. Vis. Exp. (2020).This video demonstrates the use of fluorescence lifetime imaging microscopy (FLIM) to assess polyQ protein aggregation in C. elegans neurons, comparing control and chaperone protein-deficient worms. Increased aggregation in chaperone-deficient worms promotes energy transfer between clustered fluorophores, reducing fluorescence lifetime.
FAQs
The measurement focuses on the time-dependent decay of phosphorescent emission rather than only its brightness. Because decay time is influenced by the probe’s surroundings, the resulting information is less dependent on how much probe is present or how strongly the sample was illuminated. This helps researchers interpret changes in tissue conditions more reliably across spatially varied biological samples.
Oxygen concentration can alter how long phosphorescent molecules remain in an excited state, changing the pattern of emitted-light decay. Mapping these lifetime changes therefore provides a way to visualize oxygenation within neural tissue. In living brain systems, this information can reveal regional or local changes in the tissue microenvironment that may not be apparent from spatial intensity alone.
The surrounding molecular environment can modify the decay behavior of an excited phosphorescent molecule. Interactions with nearby targets may also change the measured lifetime, allowing spatial maps to report more than probe distribution. Researchers can use these variations to investigate microenvironmental changes and examine how local conditions differ across complex neural tissues.
A lifetime change should be considered in relation to oxygen concentration, molecular environment, and interactions with nearby targets, because each can influence emission decay. Excitation intensity and probe concentration are less central to the lifetime readout than to brightness-based measurements, but the mapped result still requires attention to the biological context. This supports cautious interpretation of neural tissue changes.
A typical workflow illuminates phosphorescent molecules, records how their emitted light decays over time, and converts those decay measurements into a spatial map. Researchers then relate regional lifetime differences to oxygenation, metabolism, or other microenvironmental changes. Combining temporal decay data with tissue location allows the same experiment to connect optical measurements with localized physiology.
It is useful when researchers need spatial information about oxygenation, cellular metabolism, or changing conditions within living neural systems. The method can associate lifetime patterns with specific tissue regions rather than reporting only a general optical signal. This makes it relevant for examining brain physiology in settings where local metabolic and oxygen-related differences are important.
In neuroscience, the technique can monitor tissue changes associated with disease while preserving spatial information about where those changes occur. It can also help evaluate oxygen-sensitive probes in complex biological tissues by showing how their lifetime responses vary with local conditions. These applications connect probe behavior with brain physiology and support studies of altered neural microenvironments.