Simultaneous quantitation of Hoechst 33342 and immunofluorescence on viable cells using a fluorescence activated cell sorter

Cytometry
M R Loken

Abstract

The cytochemical stain Hoechst 33342 has been used to quantify DNA in viable cells and has been used under nonsaturating conditions to discriminate between lymphoid cell types. In order to correlate the quantitative emission from Hoechst 33342 with cell surface antigens, a fluorescence activated cell sorter was modified to simultaneously detect emission from the UV excited Hoechst dye and fluorescein attached to the cell surface by immunofluorescence techniques. A special set of laser mirrors was installed in an argon ion laser so that all the lines from 351-488 nm could be used to illuminate the cells. Appropriate emission filters were used to separate the light emitted by Hoechst 33342 from the fluorescein. An electronic cross-over circuit was used to compensate for special overlap between the two dyes. Analysis of murine lymph node cells stained both with Hoechst 33342 under nonsaturating conditions and anti-Thy 1.2 indicated that the cells that stained dimly with the Hoechst dye expressed the Thy 1.2 marker while the cells that were brightly stained with Hoechst 33342 lacked this differentiation antigen. The correlation of cell surface myeloma protein with cell cycle on an in vitro cell line indicated that the heterogeneity...Continue Reading

References

Jul 1, 1977·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·D J Arndt-Jovin, T M Jovin
Jan 1, 1979·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·M E Lalande, R G Miller
Dec 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·P J LeibsonD A Rowley
Jun 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·P J LeibsonH Schreiber
Jan 1, 1977·Immunological Reviews·H Cantor, E A Boyse
Jul 1, 1977·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·M R LokenL A Herzenberg
Jan 1, 1976·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·S A Latt, G Stetten
Aug 1, 1971·The Journal of Cell Biology·C A PasternakD B Thomas
Mar 1, 1971·Proceedings of the National Academy of Sciences of the United States of America·M Cikes, S Friberg
Jan 1, 1980·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·M R Loken
Aug 1, 1969·FEBS Letters·J J.M. BergeronC A. Pasternak

❮ Previous
Next ❯

Citations

Feb 15, 1988·Clinica Chimica Acta; International Journal of Clinical Chemistry·D H RyanP K Horan
Jan 1, 1982·Journal of Immunological Methods·M R Loken, A M Stall
Nov 16, 1984·Journal of Immunological Methods·R M Böhmer, N J King
Sep 3, 1985·Journal of Immunological Methods·R E Duerst, C N Frantz
Jan 11, 2001·Journal of Immunological Methods·I SchmidB D Jamieson
Feb 1, 1983·The Journal of Experimental Medicine·T CotnerJ Strominger
Nov 22, 2008·Journal of Biomedical Optics·Darya Yu OrlovaValeri P Maltsev
Jan 1, 1984·Cancer Investigation·F Traganos
Jan 1, 1984·Cancer Investigation·F Traganos
Sep 1, 1984·Cytometry·R E WorthingtonS Micko
Sep 1, 1986·International Journal of Cell Cloning·M P Bodger, T W LeBien
Sep 1, 1986·International Journal of Cell Cloning·P C Keng
Jan 1, 1987·Journal of Neuroscience Research·F K RenoldD G Payan
Jul 4, 2002·Hematology/oncology Clinics of North America·J Philip McCoy
Sep 15, 1982·Analytical Biochemistry·H S Kruth
Mar 14, 2013·Biochimica Et Biophysica Acta·Natasha S BartenevaIvan A Vorobjev
May 1, 1983·Immunology Today·D R ParksL A Herzenberg
Sep 5, 2008·Current Protocols in Cytometry·Z Darzynkiewicz, G Juan
Apr 2, 2011·Current Protocols in Cytometry·Zbigniew Darzynkiewicz
Apr 8, 2010·Current Protocols in Cytometry·Zbigniew Darzynkiewicz
Nov 14, 2018·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·William TelfordPascal Voluer

❮ 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.