Axioimager z2 apotome

Images from immunoperoxidase‐stained sections were captured using a Leica slide scanner and Zeiss Axioplan 2 microscope; and from immunofluorescent‐stained sections with a Zeiss Axioimager Z2 apotome. Images were adjusted for brightness and sharpness using Adobe photoshop CS6 software. Cells were counted from five sections selected at intervals along the anterior–posterior axis of each fetal sample (12 PCW, n = 2 and 19 PCW, n = 2, therefore n = 10 for each age). Sections were observed under medium magnification; rectangular counting boxes 100 μm in width were placed over the ventricular/subventricular zones (VZ/SVZ) and intermediate zone/cortical plate (IZ/CP) delineated by the nuclear staining (DAPI).

Images from immunoperoxidase stained sections were captured using a Leica slide scanner and Zeiss Axioplan 2 microscope; from immunofluorescent stained cells and sections with a Zeiss Axioimager Z2 apotome. Images were adjusted for brightness and sharpness using Adobe Photoshop CS6 software. Cells were counted from 5 sections from each fetal sample (8 PCW, n = 2 and 12 PCW, n = 2, therefore n = 10 for each age). Sections were observed under medium magnification, rectangular counting boxes of 100 μm width were placed over the ventricular/subventricular zones (VZ/SVZ) and intermediate zone/CP (IZ/CP) delineated by the nuclear staining (DAPI) on intact parts of the anterior and posterior cortex but otherwise without examining the section first. For quantification in dissociated culture, cells were cultured from 3 different fetal brains in 12-well plates, each staining combination made in duplicate. Photomicrographs were captured from 3 random fields of view from each well from which cell counts were made (therefore n = 18). Mean values with the standard error were calculated. Experimental groups were compared using a 2-tailed t-test.

Images from immunoperoxidase-stained sections were captured using a Leica slide scanner and Zeiss Axioplan 2 microscope, and from immunofluorescent-stained sections with a Zeiss Axioimager Z2 apotome. Images were adjusted for brightness and sharpness using Adobe PHOTOSHOP CS6 software. Double labeling of cells for SCGN with either DLX2 or CalR was counted from five sections selected at intervals along the anterior-posterior axis of sections from two samples aged 19 PCW. The sections were observed under medium magnification; rectangular counting boxes 100 μm in width were placed over the ventricular/subventricular zones (VZ/SVZ) and intermediate zone/cortical plate (IZ/CP) delineated by the nuclear staining (DAPI).