27 February 2007 Nanocrystal clusters in combination with spectral imaging to improve sensitivity in antibody labeling applications of fluorescent nanocrystals
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Abstract
Composition-tunable nanocrystals are fluorescent nanoparticles with a uniform particle size and with adjustable optical characteristics. When used for optical labeling of biomolecular targets these and other nanotechnology solutions have enabled new approaches which are possible because of the high optical output, narrow spectral signal, consistent quantum efficiency across a broad emission range and long lived fluorescent behavior of the nanocrystals. When coupled with spectral imaging the full potential of multiplexing multiple probes in a complex matrix can be realized. Spectral imaging can be used to improve sensitivity of narrowband fluorophores through application of chemometric image processing techniques used to reduce the influence of autofluorescence background. Composition-tunable nanocrystals can be complexed together to form nanoclusters which have the advantage of significantly stronger signal and therefore a higher sensitivity. These nanoclusters can be targeted in biomolecular systems using standard live-cell labeling and immunohistochemistry based techniques. Composition-tunable nanocrystals and nanoclusters have comparable mass and brightness across a wide emission range. This enables the production of nanocrystal-based probes that have comparable reactivity and sensitivity over a large color range. We present spectral imaging results of antibody targeted nanocrystal cluster labeling of target proteins in cultured cells and a Western blot experiment. The combination of spectral imaging with the use of clusters of nanocrystals further improves the sensitivity over either of the approaches independently.
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John S. Maier, Janice L. Panza, Matt Bootman, "Nanocrystal clusters in combination with spectral imaging to improve sensitivity in antibody labeling applications of fluorescent nanocrystals", Proc. SPIE 6448, Colloidal Quantum Dots for Biomedical Applications II, 64480P (27 February 2007); doi: 10.1117/12.701276; https://doi.org/10.1117/12.701276
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