La presentazione è in caricamento. Aspetta per favore

La presentazione è in caricamento. Aspetta per favore

Exploring synaptic circuits with antibodies and confocal microscopy PartII: Principles and applications of confocal microscopy Marco Sassoè Dip. Anatomia,

Presentazioni simili


Presentazione sul tema: "Exploring synaptic circuits with antibodies and confocal microscopy PartII: Principles and applications of confocal microscopy Marco Sassoè Dip. Anatomia,"— Transcript della presentazione:

1 Exploring synaptic circuits with antibodies and confocal microscopy PartII: Principles and applications of confocal microscopy Marco Sassoè Dip. Anatomia, Farmacologia e Medicina Legale Istituto Nazionale di Neuroscienze Torino, Italy Doctoral School of Neuroscience Turin University June 2009

2 3. In a confocal microscope the vertical resolution is: a.The same as the lateral resolution b.Better than the lateral resolution c.Worse than the lateral resolution 2. Compared to a conventional epifluorescence microscope a confocal microscope affords: a.A much better resolution b.A marginally better resolution c.The same resolution 5. In a confocal microscope the specimen is: a.Illuminated point-by-point b.Illuminated line-by-line c.Completely illuminated 1. The major advantage of a confocal microscope is: a.The use of laser illumination b.The elimination of out-of-focus fluorescent light c.The high resolution d.The presence of a Z-motor 4. The Airy Unit defines: a.The size of the pinhole b.The size of the laser beam c.The intensity of the laser beam

3

4 Development of GABAergic synapses: from synaptic adhesion to activity- dependent synaptogenesis Synaptic integration of adult-generated neurons Synapse organization in a mouse model of Rett syndrome Synaptic plasticity induced by morphine in the mesolimbic pathway Laboratory of Synapse Development and Plasticity Federica Briatore Laura Viltono Elena Frola Alessandro Ciccarelli Marta Pallotto – Patrizia Panzanelli Elena Boggio Annarita Patrizi Giulio Srubek Tomassy Maurizio Giustetto

5

6 Synapses: ( trillion) 1 mm 3 of cerebral cortex: one billion synapses!

7

8 Martin Chalfie Osamu Shimomura Roger Y. Tsien The Nobel Prize in Chemistry 2008

9

10 Leliminazione genetica di MeCP2 induce alterazioni sinaptiche neurone-specifiche VI I strato IV V II-III 100 µm WT GFP / NeuN KO WT Strato1Strato1 KO WT Strato5Strato5 Elena Boggio Maurizio Giustetto

11 Marta Pallotto WTKO Ruolo dellinibizione GABAergica nellintegrazione sinaptica dei neuroni neogenerati

12

13 Resolution R = 0,5 / NA

14 GABA GABA A R 1 PSD95 Fritschy et al., 2006 Panzanelli et al., 2004 Sassoè-Pognetto et al., 2003

15

16 Microscopio a epifluorescenza Fluorescenza excitation emission fluorochromes fluorophores autofluorescence Indirect (secondary) fluorescence

17

18 The pinhole is conjugate to the focal point of the lens Sistema confocale Sorgente luce puntiforme Illuminazione puntiforme del campione (con obiettivo ad elevata AN) Pinhole su piano di rilevamento Vantaggi: -Eliminazione piani fuori fuoco – immagini più nitide -Eliminazione di interferenza laterale – aumento contrasto -Sezioni ottiche (e ricostruzioni 3D) -Miglior risoluzione (teorico) TUBE LENS confocal


Scaricare ppt "Exploring synaptic circuits with antibodies and confocal microscopy PartII: Principles and applications of confocal microscopy Marco Sassoè Dip. Anatomia,"

Presentazioni simili


Annunci Google