Lsm 510 pascal

Samples were fixed in the formaldehyde vapour during an hour and stained with calcofluor (excitation 405 nm, filter BP 420-480), ethidium bromide (Sigma, USA) (excitation 514 nm, filter BP 530-600 nm), and thiazine dyes (excitation 514 nm, filter BP 530-600 nm). A microscopic examination of sample fluorescence was performed, using CSLM AXIOSKOP -2 ZEISS equipped by the LSM 510 PASCAL (CarlZeiss, FRG) software.

Morphological analyses were conducted with a LSM510 Pascal (Zeiss) confocal scanning microscope. Optical sections were acquired at 1.0-1.3-mm intervals and reconstructed three-dimensionally with Amira software ver. 5.5 (Visage Imaging GmbH). Selected neuronal regions were further processed using surface and volume rendering functions to calculate the volume of the region of interest and generate 3D reconstructions. The terminal varicosities were readily identifiable from the characteristic swellings located around the tips of axonal branches [18] .
The distribution pattern of PNs' dendritic terminals within the central region of the glomerulus (20 mm in thickness, corresponding to 15-16 optical sections) was detected using Amira software, and further processed by means of a custom-written Excel macro (Microsoft, 2010 version) for glomerular size normalization. The shortest distance from the lateral half outline of each glomerulus was then quantified.

Confocal Scanning Laser Microscopy (CSLM) Samples were fixed in the formaldehyde vapor during an hour and stained with calcofluor (excitation 405 nm, filter BP 420-480), and thiazine dyes (excitation 514 nm, filter BP 530-600 nm). A microscopic examination of sample fluorescence was performed, using CSLM AXIOSKOP -2 ZEISS equipped by the LSM 510 PASCAL (CarlZeiss, FRG) software.