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Optics
Focus
Confocal theory
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Dr. Marvin MinskyConfocal microscope: 1957
Confocal theory
LASER
LightAmplification by theStimulatedEmission ofRadiation
ScanningConfocalMicroscope
Laser
DichroicMirror
Conventional / Confocal
Fluorescence
400 nm
Wavelength and Light Properties
575 nm
E = h c
HighEnergy
LowEnergy0,5*
*2
Multiphoton exitation
800 nm
800 nm
1200 nm
1200 nm
1200 nm
400 nm
Dr. Maria Göppert-Mayer : theory of two-photon quantum transitions (two-photon absorption and emission) 1931,
LASER500 nm
Pulsed LASER1000 nm1200 nm800 nm
nm500 nm800 1200
Temporal photon concentration
Prof. Watt W. Webb et al.Two-photon laser scanning fluorescence microscopy:
1990
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Spatial photon concentration
Confocal / Multiphoton
Luo at al. Cell Structure and Function 2006, 31: 63Comparison of photoactivation of PA-GFP in vivo with single-photon (405 nm) and multiphoton (790 nm) laser light.
PhotoconversionExcitation area
Thorsten at al. Nature 2004; 427: 159T-cell priming by dendriticcells in lymph nodes occurs in three distinct phases
In VIVO Imaging
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