Correlative Confocal and 3D Electron Microscopy of a Specific Sensory Cell.
Delineation of a cell's ultrastructure is important for understanding its function. This can be a daunting project for rare cell types diffused throughout tissues made of diverse cell types, such as enteroendocrine cells of the intestinal epithelium. These gastrointestinal sensors of food and bacteria have been difficult to study because they are dispersed among other epithelial cells at a ratio of 1:1,000. Recently, transgenic reporter mice have been generated to identify enteroendocrine cells by means of fluorescence. One of those is the peptide YY-GFP mouse. Using this mouse, we developed a method to correlate confocal and serial block-face scanning electron microscopy. We named the method cocem3D and applied it to identify a specific enteroendocrine cell in tissue and unveil the cell's ultrastructure in 3D. The resolution of cocem3D is sufficient to identify organelles as small as secretory vesicles and to distinguish cell membranes for volume rendering. Cocem3D can be easily adapted to study the 3D ultrastructure of other specific cell types in their native tissue.
Department of Medicine, Duke University Medical Center; diego.bohorquez@duke.edu.
Diego
D
Bohórquez
Department of Chemistry, University of North Carolina - Chapel Hill.
Fariha
F
Haque
Renovo Neural Incorporated.
Satish
S
Medicetty
Department of Medicine, Duke University Medical Center.
Rodger A
RA
Liddle
D000818:Animals; D019858:Enteroendocrine Cells / Q000166:cytology / Q000000981:diagnostic imaging*; D021621:Imaging, Three-Dimensional / Q000379:methods; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal / Q000379:methods*; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D014463:Ultrasonography