Out of the blue: a comparison of Hoechst side population (SP) analysis of murine bone marrow using 325, 363 and 407 nm excitation sources

Journal of Immunological Methods
Catherine SimpsonDerek Davies

Abstract

The discrimination of side population (SP) cells is a useful flow cytometric tool for the study of putative stem cells in many tissue types. Previous studies on murine bone marrow (BM) SP cells and concurrent surface immunophenotyping provide a good model for comparing Hoechst bivariate profiles. Here we compare the results of SP cells from the same specimens generated on three different instruments. Concurrent SP analysis and immunophenotyping was performed on murine BM. The data acquisition was performed on a DakoCytomation MoFlo, a Becton Dickinson LSR and a Becton Dickinson FACSAria to compare the ability of each instrument to discriminate putative stem cells. Further experiments examined the effect of apoptotic changes, assessed by Annexin V binding, and the presence of nucleated erythroid precursors on the Hoechst profile. Identifiable and equivalent side populations were generated on all three instruments despite differences in appearance of the Hoechst profiles. Differences in laser type, collection filter combinations and the presence of erythroid precursors and apoptotic cells all contributed to this effect. In our hands, murine SP cells with the expected surface immunophenotype can be identified on a MoFlo, LSR and a...Continue Reading

References

Aug 30, 1992·Journal of Immunological Methods·M G OrmerodD Robertson
Apr 1, 1996·The Journal of Experimental Medicine·M A GoodellR C Mulligan
May 30, 2001·Cytometry·H M Shapiro, N G Perlmutter
May 27, 2003·Investigative Ophthalmology & Visual Science·Sumitra BhattacharyaIqbal Ahmad
Jul 25, 2003·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Rupesh I BhattNoel W Clarke
Dec 3, 2003·Blood·Daniel J PearceDominique Bonnet
Dec 31, 2003·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·William G Telford, Ella G Frolova
Aug 4, 2004·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Timothy W PetersenGer van den Engh

❮ Previous
Next ❯

Citations

Dec 23, 2006·Stem Cells·William G TelfordSusan E Bates
Dec 13, 2006·Methods in Enzymology·K K Lin, Margaret A Goodell

❮ Previous
Next ❯

Related Concepts

Related Feeds

Anthelmintics

Anthelmintics or antihelminthics are a group of antiparasitic drugs that expel parasitic worms (helminths) and other internal parasites from the body by either stunning or killing them and without causing significant damage to the host. Discover the latest research on anthelmintics here.

Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis