Imaging spatiotemporal dynamics of neuronal signaling using fluorescence resonance energy transfer and fluorescence lifetime imaging microscopy

  title={Imaging spatiotemporal dynamics of neuronal signaling using fluorescence resonance energy transfer and fluorescence lifetime imaging microscopy},
  author={Ryohei Yasuda},
  journal={Current Opinion in Neurobiology},
  • R. Yasuda
  • Published 1 October 2006
  • Biology, Chemistry, Physics
  • Current Opinion in Neurobiology

Principle and Application of Fluorescence Lifetime Imaging for Neuroscience

  • R. Yasuda
  • Biology, Chemistry
    Neurophotonics and Biomedical Spectroscopy
  • 2019

Optogenetic Imaging of Protein Activity Using Two-Photon Fluorescence Lifetime Imaging Microscopy.

  • H. Murakoshi
  • Biology, Chemistry
    Advances in experimental medicine and biology
  • 2021
Of the various FRET detection systems, two-photon fluorescence lifetime imaging microscopy (2pFLIM) is one of the methods best suited to monitoring FRET in subcellular compartments of living cells located deep within tissues, such as brain slices.

Optogenetic Imaging of Protein Activity in the Synapse Using 2-Photon Fluorescence Lifetime Imaging Microscopy

The principle of the 2pFLIM-FRET is introduced for monitoring intracellular protein activities and protein–protein interactions using two examples: detecting small GTPase activity and monitoring actin polymerization in dendrites and synapses of hippocampal neurons in brain slices.

Imaging the activity of Ras superfamily GTPase proteins in small subcellular compartments in neurons.

This protocol describes the procedures and equipment required for imaging intracellular signaling activity, with a particular focus on signaling mediated by the Ras superfamily of small GTPase proteins.

Two-Photon Lifetime Imaging of Signal Transduction in Neurons

Recent results of the imaging of CamKII and Ras activation in single dendritic spines in brain tissue are introduced and 2-photon Fluorescence Lifetime Imaging Microscopy (2pFLIM) is introduced.

Fluorescence of Strongly Absorbing Multicomponent Media

All characteristics of fluorescence – intensity, spectrum, polarization, lifetime and quantum yield – are used now for research and diagnostics in biology and medicine.

Imaging Neuromodulatory Signaling Events at Single Cell Resolution in Behaving Animal

The work in the development of genetically encoded indicators for monitoring intracellular events downstream of neuromodulation, and implementing in vivo imaging of these indicators in combination with two-photon fluorescent lifetime imaging microscopy (2pFLIM) is presented.

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy.

In vivo PKA imaging with single-cell resolution was developed as a readout for neuromodulatory events in a manner analogous to calcium imaging for neuronal electrical activities, and will be applicable to many other brain regions to study corresponding behavior-induced PKA activities.

Fluorescent sensors for neuronal signaling




Imaging protein molecules using FRET and FLIM microscopy.

Fluorescence lifetime imaging for the two-photon microscope: time-domain and frequency-domain methods.

A comparison of the performance of the two approaches in terms of signal-to-noise ratio (SNR) and the speed of data acquisition shows the frequency-domain method is faster and provides less distortion for bright samples.

Multiphoton-FLIM quantification of the EGFP-mRFP1 FRET pair for localization of membrane receptor-kinase interactions.

An improved monomeric form of the red fluorescent protein, mRFP1, is presented as the acceptor in biological fluorescence resonance energy transfer (FRET) experiments using the enhanced green fluorescent protein as donor and should be widely applicable, particularly to live-cell quantitative FRET assays.

Development of a multiphoton fluorescence lifetime imaging microscopy system using a streak camera

We report the development and detailed calibration of a multiphoton fluorescence lifetime imaging system (FLIM) using a streak camera. The present system is versatile with high spatial (∼0.2 μm) and

Supersensitive Ras activation in dendrites and spines revealed by two-photon fluorescence lifetime imaging

It is found in CA1 hippocampal neurons that trains of back-propagating action potentials rapidly and reversibly activated Ras in dendrites and spines, and the Ras pathway functions as a supersensitive threshold detector for neural activity and Ca2+ concentration.

Multiple Time-Gate Module for Fluorescence Lifetime Imaging

A versatile multiple time-gate fluorescence lifetime acquisition module is presented. The module is used as an add-on to a two-photon scanning laser microscope equipped with pulsed excitation for

Quantitative imaging of lateral ErbB1 receptor signal propagation in the plasma membrane.

The rapid and extensive propagation of receptor phosphorylation over the entire cell after focal stimulation demonstrates a signaling wave at the plasma membrane resulting in full activation of all receptors.