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Proceedings Paper

Nanocrystal-based biomimetic system for quantitative flow cytometry
Author(s): Peter Yim; Marina Dobrovolskaia; HyeongGon Kang; Matthew Clarke; Anil K. Patri; Jeeseong Hwang
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Paper Abstract

Flow cytometry has been instrumental in rapid analysis of single cells since the 1970s. One of the common approaches is the immunofluorescence study involving labeling of cells with antibodies conjugated to organic fluorophores. More recently, as the application of flow cytometry extended from simple cell detection to single-cell proteomic analysis, the need of determining the actual number of antigens in a single cell has driven the flow cytomery technique towards a quantitative methodology. However, organic fluorophores are challenging to use as probes for quantitative detection due to the lack of photostability and of quantitative fluorescence standards. National Institute of Standards and Technologies (NIST) provides a set of fluorescein isothiocyanate (FITC) labeled beads, RM 8640, which is the only nationally recognized fluorescent particle standard. On the other hand, optical characteristics of semiconductor nanocrystals or quantum dots or QDs are superior to traditional dye molecules for the use as tags for biological and chemical fluorescent sensors and detectors. Compelling advantages of QDs include long photostability, broad spectral coverage, easy excitation, and suitability for multiplexed sensing. Recently, novel surface coatings have been developed to render QDs water soluble and bio-conjugation ready, leading to their use as fluorescent tags and sensors for a variety of biological applications including immunolabeling of cells. Here, we describe our approach of using fluorescent semiconductor QDs as a novel tool for quantitative flow cytometry detection. Our strategy involves the development of immuno-labeled QD-conjugated silica beads as "biomimetic cells." In addition to flow cytometry, the QD-conjugated silica beads were characterized by fluorescence microscopy to quantitate the number of QDs attached to a single silica bead. Our approach enables flow cytometry analysis to be highly sensitive, quantitative, and encompass a wide dynamic range of fluorescence detection. Quantitative aspects of the proposed flow cytometery-based approach for measurement of the QD-based biomimetic samples are discussed.

Paper Details

Date Published: 6 February 2007
PDF: 9 pages
Proc. SPIE 6430, Advanced Biomedical and Clinical Diagnostic Systems V, 64301T (6 February 2007); doi: 10.1117/12.714138
Show Author Affiliations
Peter Yim, National Institute of Standards and Technology (United States)
Marina Dobrovolskaia, SAIC-Frederick/NCI-Frederick (United States)
HyeongGon Kang, National Institute of Standards and Technology (United States)
Matthew Clarke, National Institute of Standards and Technology (United States)
Anil K. Patri, SAIC-Frederick/NCI-Frederick (United States)
Jeeseong Hwang, National Institute of Standards and Technology (United States)

Published in SPIE Proceedings Vol. 6430:
Advanced Biomedical and Clinical Diagnostic Systems V
Ramesh Raghavachari; Tuan Vo-Dinh; Warren S. Grundfest M.D.; David A. Benaron M.D.; Gerald E. Cohn, Editor(s)

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