Leica dfc9000 gtc vsc 09991 camera

A randomly chosen series was used to calculate the total volume of the DG in each animal using Nissl staining. Slices were mounted on 2% gelatine-coated glass slides and air-dried at rt for 48 h. The slides were sequentially immersed in the following solutions: 6 min in toluidine blue, 10 sec in bi-distilled water, 2 min in EtOH 70°, 2 min in EtOH 96°, 2 min in EtOH 100°, 2 min in EtOH 100°, and 2 min in Xylene (PanReac, #251769). Next, sections were embedded in DePex (Serva Electrophoresis™, #1824301) and dried for 48 h at rt before imaging. Images were acquired under a THUNDER Imager Tissue microscope equipped with a Leica DFC9000 GTC VSC-09991 camera (Leica Microsystems Ltd., Wetzlar, Germany) and using a 5X dry objective. Images were processed using the Leica Application Suite X (LAS X) software provided by the manufacturer (Leica Microsystems Ltd., Wetzlar, Germany). The DG volume was determined using the freehand selection tool in Fiji software to measure the granule cell layer (GCL) plus the SGZ area on each section of the series. Each area was multiplied by the thickness of the tissue (namely 50 µm) and by the sampling fraction (8). The numbers obtained were summed to calculate the total DG volume, which is expressed in mm³.

Images were acquired under a THUNDER Imager Tissue microscope, equipped with a Leica DFC9000 GTC VSC-09991 camera (Leica Microsystems Ltd., Wetzlar, Germany). For Extended Depth of Field (EDF) images containing the whole hippocampus, an HC PL APO 20x/0.80 DRY objective and Tile Scan acquisition mode were used. For representative images of neuronal morphology, an HC PL APO 40x/0.95 and a 63x/0.95 oil objective lens were used. Images were processed using the Leica Application Suite X (LAS X) software provided by the manufacturer (Leica Microsystems Ltd., Wetzlar, Germany).