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An Innovative Triple Immunogold Labeling Method to Investigate the Hemopoietic Stem Cell Niche In Situ

Published online by Cambridge University Press:  16 September 2009

Sarah L. Ellis ,
Brenda Williams ,
Stephen Asquith ,
Ivan Bertoncello  and
Susan K. Nilsson
Sarah L. Ellis
Affiliation:
Microscopy Core Facility, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia Australian Stem Cell Centre, Clayton, Victoria 3168, Australia Department of Pathology, University of Melbourne, Parkville, Victoria 3052, Australia
Brenda Williams
Affiliation:
Australian Stem Cell Centre, Clayton, Victoria 3168, Australia
Stephen Asquith
Affiliation:
Microscopy Core Facility, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia
Ivan Bertoncello
Affiliation:
Australian Stem Cell Centre, Clayton, Victoria 3168, Australia Department of Pathology, University of Melbourne, Parkville, Victoria 3052, Australia
Susan K. Nilsson*
Affiliation:
Australian Stem Cell Centre, Clayton, Victoria 3168, Australia Department of Pathology, University of Melbourne, Parkville, Victoria 3052, Australia
*
Corresponding author. E-mail: [email protected]
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Abstract

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.

Keywords

hemopoietic stem cellhemopoietic progenitor cellhomingnicheimmunogoldhyaluronic acidosteopontin
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 15 , Issue 5 , October 2009 , pp. 403 - 414
Copyright
Copyright © Microscopy Society of America 2009

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References

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