Detection and isolation of rare cells by 2-step enrichment high-speed flow cytometry/cell sorting and single cell LEAP laser ablation

M. D. Zordan; James F. Leary

doi:10.1117/12.874270

11 February 2011 Detection and isolation of rare cells by 2-step enrichment high-speed flow cytometry/cell sorting and single cell LEAP laser ablation

M. D. Zordan, James F. Leary

Author Affiliations +

Proceedings Volume 7902, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IX; 790213 (2011) https://doi.org/10.1117/12.874270
Event: SPIE BiOS, 2011, San Francisco, California, United States

Abstract

The clonal isolation of rare cells, especially cancer and stem cells, in a population is important to the development of improved medical treatment. We have demonstrated that the Laser-Enabled Analysis and Processing (LEAP, Cyntellect Inc., San Diego, CA) instrument can be used to efficiently produce single cell clones by photoablative dilution. Additionally, we have also shown that cells present at low frequencies can be cloned by photoablative dilution after they are pre-enriched by flow cytometry based cell sorting. Circulating tumor cells were modeled by spiking isolated peripheral blood cells with cells from the lung carcinoma cell line A549. Flow cytometry based cell sorting was used to perform an enrichment sort of A549 cells directly into a 384 well plate. Photoablative dilution was performed with the LEAP^TM instrument to remove any contaminating cells, and clonally isolate 1 side population cell per well. We were able to isolate and grow single clones of side population cells using this method at greater than 90% efficiency. We have developed a 2 step method that is able to perform the clonal isolation of rare cells based on a medically relevant functional phenotype.

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M. D. Zordan and James F. Leary "Detection and isolation of rare cells by 2-step enrichment high-speed flow cytometry/cell sorting and single cell LEAP laser ablation", Proc. SPIE 7902, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IX, 790213 (11 February 2011); https://doi.org/10.1117/12.874270

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KEYWORDS

Blood

Luminescence

Biological research

Flow cytometry

Laser ablation

Electronic filtering

Analytical research

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