An innovative triple immunogold labeling method to investigate the hemopoietic stem cell niche in situ.
The ultrastructural study of rare cells within their niche in situ is very difficult. We have developed a method for locating individual transplanted cells and simultaneously identifying and analyzing the molecules and cellular phenotypes surrounding them in situ using transmission electron microscopy. This innovative method involves triple immunogold labeling combined with serial ultrathin sectioning. We demonstrate the validity of this approach by examining the niche of individual transplanted cells from a population highly enriched for hemopoietic stem cells and the ultrastructural expression of two key stem cell regulatory molecules, hyaluronic acid and osteopontin. In addition, we describe the phenotypes of the surrounding cells.
Ellis
Sarah L
SL
Microscopy Core Facility, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia.
Williams
Brenda
B
Asquith
Stephen
S
Bertoncello
Ivan
I
Nilsson
Susan K
SK
D000818:Animals; D006412:Hematopoietic Stem Cells / Q000737:chemistry* / Q000648:ultrastructure*; D006820:Hyaluronic Acid / Q000032:analysis; D007150:Immunohistochemistry / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D053495:Osteopontin / Q000032:analysis; D013194:Staining and Labeling / Q000379:methods*; D055153:Stem Cell Niche
D053495:Osteopontin; D006820:Hyaluronic Acid