Epifluorescence Confocal. Zoom 1 Zoom 2 Zoom 4.

Epifluorescence

Confocal

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Point Spread Function

x - y x - z

Disco di Airy

3D Deconvolution

Sassoè-Pognetto et al., J Comp Neurol 2000

Sassoè-Pognetto et al., J Comp Neurol 2000

FITC-TRITC emission

WT GABAAR1 KOVGAT/Calb

Wulff et al., Nat Neurosci in press

GABA-positive terminals in the molecular layermake asymmetric synapses with Purkinje cell spines

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GABAGABA

parallel fiberparallel fiber

Fritschy et al., J Neurosci 2006

sp

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sp

sp

NL2 does not localize ectopically at heterologous synapses with PC spines

GABAGABA

NL2/mGluR1/GAD

NL2/Car8/GAD

Patrizi et al., PNAS 2008

Schneider Gasser et al et al.,

Two-photon microscopy

Two photons of low energy (infrared range) can excite a fluorophore → emission of a fluorescence photon at a higher energy

Excitation is restricted to a tiny focal volume (rejection of out-of-focus signals)

Longer wavelengths penetrate deeper into tissue(1 mm)

Lower energy photons cause relatively low damage outside the focal volume → in vivo imaging