Journal: Developmental neurobiology

Article Title: Glial Cell Expansion Coincides with Neural Circuit Formation in the Developing Auditory Brainstem

doi: 10.1002/dneu.22633

Figure Lengend Snippet: Experimental design for cell counting using 3D virtual reality. (A) Serial sections containing MNTB from Cre;Ai9 mice were stained with cell-specific antibodies for neurons (Map2), astrocytes (Aldh1L1), oligodendrocytes (Sox10) and microglia (Iba1), in an alternating manner, along with DAPI. These slices were imaged with a 40× objective and tiled. The MNTB is outlined in white in each section and the midline is indicated by a dashed white line. Scale bar = 100 μm. (B) The MNTB was cropped from the larger image and imported into syGlass for cell counting in 3D virtual reality. The solid and dashed white lines demarcate cut planes that were rotated to generate the 3D view in C (arrow). Scale bar = 50 μm. (C) Rotated view of part of MNTB to illustrate 3D view for cell counting. Counting dots were placed onto the nuclei. White dots indicate incomplete (sectioned) nuclei in the first slice of the z-stack. These were excluded from analysis, but incomplete nuclei in the bottom slice were counted and included in the analysis. Red dots indicate cells that were counted. Note that cells throughout the entire MNTB (panel B) were counted.

Article Snippet: The syGlass 3D immersive visualization program was used to perform cell counts because it enables rapid and accurate assessment of sectioned DAPI-labeled nuclei at the edge of the first image plane , and it permits accurate assessment of spatial association of markers with DAPI in three dimensions.

Techniques: Cell Counting, Staining