A nouvel method to visualize tissue molecular architecture at the level of synapse

• Autorzy: Szepesi Z. , Szczepankiewicz A. , Walczak A. , Wilczynski G. , Wlodarczyk J. , Kaczmarek L.
• Języki publikacji: EN

Abstrakty

EN

To understand various biological functions of CNS, visualization of the fi ne details of tissue molecular architectures and synaptic structures is of the major importance. Herein, we use a method for immunfl uorescence microscopy called Array Tomography which allows for imaging details of tissue structure. To this end, we cut resin-embedded specimen into serial sections (50–250 nm) by ultramicrotome. Hydrophilic resin permits the antibody penetration into the tissue, thus providing an opportunity for post-embedding immunostaining. The ribbon of serial sections represents tissue volume. Reconstruction of three dimensional distributions of antigens is obtained by imaging and post-processing of consecutive images. The array can be eluted and re-stained, thus allowing for detection of several antigens in the same sample. The above methodology provides visualization at the spatial resolution that is higher than the one obtained using conventional confocal microscopy. Furthermore, we use this method together with the super-resolution (70 nm) stimulated emission depletion (STED) microscopy. Thus, combining of these two methods provides very detailed image of tissue architecture and offers a novel opportunity to reconstruct the molecular architecture with sub-micrometer resolution in three dimensions.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2009
• Tom: 69
• Numer: 3
• Opis fizyczny: p.319

Twórcy

autor

Szepesi Z.

• Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland

autor

Szczepankiewicz A.

• Department of Neurophysiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland

autor

Walczak A.

• Department of Neurophysiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland

autor

Wilczynski G.

• Department of Neurophysiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland

autor

Wlodarczyk J.

• Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland

autor

Kaczmarek L.

• Department of Molecular and Cellular Neurobiology, Nencki Institute of Experimental Biology Polish Academy of Sciences, Warsaw, Poland