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Proceedings Paper

Immunomagnetic cell separation, imaging, and analysis using Captivate ferrofluids
Author(s): Laurie Jones; Joseph M. Beechem
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Paper Abstract

We have developed applications of CaptivateTM ferrofluids, paramagnetic particles (approximately 200 nm diameter), for isolating and analyzing cell populations in combination with fluorescence-based techniques. Using a microscope-mounted magnetic yoke and sample insertion chamber, fluorescent images of magnetically captured cells were obtained in culture media, buffer, or whole blood, while non-magnetically labeled cells sedimented to the bottom of the chamber. We combined this immunomagnetic cell separation and imaging technique with fluorescent staining, spectroscopy, and analysis to evaluate cell surface receptor-containing subpopulations, live/dead cell ratios, apoptotic/dead cell ratios, etc. The acquired images were analyzed using multi-color parameters, as produced by nucleic acid staining, esterase activity, or antibody labeling. In addition, the immunomagnetically separated cell fractions were assessed through microplate analysis using the CyQUANT Cell Proliferation Assay. These methods should provide an inexpensive alternative to some flow cytometric measurements. The binding capacities of the streptavidin- labled Captivate ferrofluid (SA-FF) particles were determined to be 8.8 nmol biotin/mg SA-FF, using biotin-4- fluorescein, and > 106 cells/mg SA-FF, using several cell types labeled with biotinylated probes. For goat anti- mouse IgG-labeled ferrofluids (GAM-FF), binding capacities were established to be approximately 0.2 - 7.5 nmol protein/mg GAM-FF using fluorescent conjugates of antibodies, protein G, and protein A.

Paper Details

Date Published: 28 May 2002
PDF: 10 pages
Proc. SPIE 4622, Optical Diagnostics of Living Cells V, (28 May 2002); doi: 10.1117/12.468339
Show Author Affiliations
Laurie Jones, Molecular Probes, Inc. (United States)
Joseph M. Beechem, Molecular Probes, Inc. (United States)

Published in SPIE Proceedings Vol. 4622:
Optical Diagnostics of Living Cells V
Robert C. Leif; Daniel L. Farkas; Robert C. Leif, Editor(s)

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