Loading presentation...

Present Remotely

Send the link below via email or IM

Copy

Present to your audience

Start remote presentation

  • Invited audience members will follow you as you navigate and present
  • People invited to a presentation do not need a Prezi account
  • This link expires 10 minutes after you close the presentation
  • A maximum of 30 users can follow your presentation
  • Learn more about this feature in our knowledge base article

Do you really want to delete this prezi?

Neither you, nor the coeditors you shared it with will be able to recover it again.

DeleteCancel

Fluorescence Lifetime Imaging Microscopy

No description
by

Paulien Stegehuis

on 9 September 2013

Comments (0)

Please log in to add your comment.

Report abuse

Transcript of Fluorescence Lifetime Imaging Microscopy

Fluorescence Lifetime Imaging Microscopy
Introduction
Fluorescence
Fluorescence lifetime
Measurements
Applications
Experiments LUMC
Setup
Endogenous
Extrinsic
Genetically
Engineered
Emission spectrum
Polarization
Quantum yield
Lifetime
Excitation spectrum
Topics
Average time a molecule stays in its excited state before it emits a photon.
Definition of FLIM
Lifetime measurements
Different applications
FLIM experiments
Time domain vs frequency domain
Wide field vs point scanning
Time domain versus Frequency domain
Questions?
Г = radiative decay
k = non-radiative decay
τ= 1/(Γ +k )
τ = fluorescence lifetime
I(t)= I(0)e^(-t/τ) + c
Lifetime
Does not depend on intensity
Depends on the environment
differentiate fluorophores with similar spectra
Point scanning versus wide field FLIM
High frequency pulsed laser
Single photons are detected
Arrival times detected

Modulated light wave
2 Parameters measured:
Phase shift & amplitude reduction
Autofluorescence
Cancer applications
For discrimination of different fractions of the same fluorophore in different states of interaction with its environment
FRET
Förster resonance energy transfer
Changes lifetime donor
More robust than intensity
pH and ion concentration
FLIM to obtain information on the environment
Christoph Biskup, University Jena, Germany
Theodora Mauro, University of San Fransisco, USA
Thomas Gensch, Julich Research Centre, Germany
Skin - lifetime as
indicator of pH
Spinal ganglion - lifetime as indicator of Cl- concentration
Paul French, Imperial College London, UK
Slow versus Fast
High sensitivity versus Low sensitivity
Higher temporal resolution versus Lower temporal resolution
Determine expected fluorescence lifetime changes in tumor specific probes.
Lifetime probe:
- Without cells
- With targeted cells
- With already saturated targeted cells
- With cells that show no/a low target expression
Full transcript