Immunomagnetic cell separation, imaging, and analysis using Captivate ferrofluids

Laurie Jones; Joseph M. Beechem

doi:10.1117/12.468339

28 May 2002 Immunomagnetic cell separation, imaging, and analysis using Captivate ferrofluids

Laurie Jones, Joseph M. Beechem

Proceedings Volume 4622, Optical Diagnostics of Living Cells V; (2002) https://doi.org/10.1117/12.468339
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

Abstract

We have developed applications of Captivate^TM 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.

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Laurie Jones and Joseph M. Beechem "Immunomagnetic cell separation, imaging, and analysis using Captivate ferrofluids", Proc. SPIE 4622, Optical Diagnostics of Living Cells V, (28 May 2002); https://doi.org/10.1117/12.468339

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KEYWORDS

Magnetism

Luminescence

Proteins

Particles

Image analysis

Microscopes

Biological research

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