Phagocytosis of Neural Debris by Cultured Glial Cells

To measure phagocytic capacity of glial cells, Dr. Won-suk Chung, a postdoctoral fellow in the Ben Barres lab, collected z-stack images of cultured, purified mammalian glial cells incubated with neural debris using the 63×/1.4 NA Plan Apochromat objective on NMS's LSM510 Meta confocal.

Panel A shows the glial cells (red) and neural debris (green—phagocytosed debris tends to appear yellow). Scale bar = 10 μm. Imaris 3D analysis software was used to quantify phagocytosis in a three-step process (B–D).

  1. Panel B: cell surfaces were defined using smoothing and thresholding operations.
  2. Panel C–D: volume of neural debris was quantified by segmenting the green channel (C) into discrete spheres (D, in blue), using intensity thresholding and size criteria.
  3. Panel E: selection of debris spots that were either touching or enclosed by the cell surface (in orange).

phagocytosis of neural debris by cultured mammalian glial cells: image data from Dr. Won-suk Chung
Image data collected on NMS's LSM510 Meta confocal by Dr. Won-suk Chung.
Analysis performed with Imaris software on NMS's CBIS-funded image analysis workstation.