Toward the Accurate Read‐out of Quantum Dot Barcodes: Design of Deconvolution Algorithms and Assessment of Fluorescence Signals in Buffer

Lee, J. A.; Hung, A.; Mardyani, Sawitri; Rhee, Alex; Klostranec, Jesse M.; Mu, Ying; Li, D.; Chan, Warren C. W.

doi:10.1002/adma.200701955

Cited by 70 publications

(81 citation statements)

References 22 publications

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“…The degradation of the QDs leads to loss of fluorescence and broadening of the emission spectra. [22] To quantify QD metabolism, we measured Cd 2þ in culture media with ICP-MS after filtering the media with 10 kDa membrane to remove any intact QDs. Mancini et al…”

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confidence: 99%

Exploring Primary Liver Macrophages for Studying Quantum Dot Interactions with Biological Systems

et al. 2010

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confidence: 99%

Exploring Primary Liver Macrophages for Studying Quantum Dot Interactions with Biological Systems

et al. 2010

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“…Despite such effects, the final barcode has a unique and recognizable signature that can be identified with a deconvolution algorithm that takes into account the relative QD concentration for each color, the photoluminescence overlapped with QD absorption profiles, photoluminescence quantum yields, and the microbead volume. [25] The most critical property of a spectroscopic barcode, however, is its robustness against changing external environments.…”

Section: Methodsmentioning

confidence: 99%

“…[20,25,28] This variability would prohibit the correct identification of the barcodes after their use in biological assays. To compare and test barcode stability, photoluminescence intensity profiles were monitored against the entire range of pH values (pH 0-14), high temperatures up to 95 8C (just below the glass transition temperature of pure polystyrene at 95 8C), and a variety of different chemical environments known to affect QD fluorescence efficiencies.…”

Section: Methodsmentioning

confidence: 99%

“…[21,22] Microbead "swelling" and layer-by-layer techniques result only in surface-level loading of QDs into the polymer, [18,23] which are thus exposed to pH values and environmental factors that destabilize their fluorescence intensity. [24,25] Stability of the barcode (fluorescence profile) requires that the QDs be positioned well within the polymer matrix and do not leak from the bead, and thus, techniques to encapsulate the QDs during the polymerization step were developed. [21,22] However, this process is lengthy, requires considerable presynthetic QD surface modification, and yet results in broadly dispersed microbead sizes.…”

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Facile and Rapid One‐Step Mass Preparation of Quantum‐Dot Barcodes

Fournier‐Bidoz¹,

Jennings²,

Klostranec³

et al. 2008

Angew Chem Int Ed

132

163

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Since their creation in 1949 by Woodland and Silver for grocery and warehouse inventory, [1] the broad application and utility of barcodes has continuously expanded. Today, in response to the demand for high-throughput multiplexed detection for elucidation of biomolecular mechanisms and to advancing personalized molecular diagnostics and therapeutics, molecular barcodes are much needed for inventorying biological molecules. [2][3][4] Within the context of molecular barcoding, two platforms capable of fulfilling the needs for code recognition and multiplexed detection exist: graphical barcodes utilizing structural recognition and spectroscopic barcodes using the unique optical properties of an embedded material. [2] Whereas each platform maintains the potential to construct large barcode libraries, limitations in barcode stability, reproducibility, or readout impose serious limitations. Graphical barcodes, [5][6][7][8][9][10][11] such as etched polymeric or striped metallic structures, suffer a multitude of problems from the complex instrumentation required for both synthesis and readout, the slow data collection rate (approximately 3 Hz for recent polymeric structures [11] compared to several kilohertz detection of microbeads by flow cytometry), to the unstable dispersion of these structures in buffer or media, making them unfit for mainstream applications. While spectroscopic barcodes employ a colorimetric readout, fluorescence-based barcodes are rapidly detected using flow cytometry [12,13] or miniaturized home-built systems.[14] Raman barcodes still require planar array readout after lengthy detection protocols.[15] From a time-efficiency and sensitivity standpoint, fluorescence-based microbead barcodes are therefore best suited for applications in high-throughput multiplexed detection.Fluorescent quantum dots (QDs) boast narrow Gaussian emission line shapes, resistance to photobleaching, high quantum yields, and single-wavelength excitation, whereas molecular dyes suffer from both photobleaching and redtailed emission; [16,17] thus, QDs are ideal fluorophores for barcoding. However, the construction of QD barcodes has shown minimal advance since their conception in 2001 [18] because of difficulties encountered in the mass production of robust and reproducible barcoded materials. Current methods to prepare QD barcodes include the "swelling" technique, [18] QD entrapment inside layer-by-layer charged polymer coatings [19] or mesoporous silica microbeads, [20] and polymerizable QD encapsulation. [21,22] Microbead "swelling" and layer-by-layer techniques result only in surface-level loading of QDs into the polymer, [18,23] which are thus exposed to pH values and environmental factors that destabilize their fluorescence intensity. [24,25] Stability of the barcode (fluorescence profile) requires that the QDs be positioned well within the polymer matrix and do not leak from the bead, and thus, techniques to encapsulate the QDs during the polymerization step were developed. [21,22] However, this process is lengthy, r...

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“…So far, both organic [7,8] and inorganic [9,10] fluorophores have been used to optically encode microspheres. However, the application of organic fluorophores in multicolor analysis is limited to problems including narrow excitation bands and broad emission spectra, which can make detection of multiple light emitting probes difficult due to spectral overlap and photodegradation [11].…”

Section: Introductionmentioning

confidence: 99%

Preparation of quantum dots encoded microspheres by electrospray for the detection of biomolecules

Sun

et al. 2011

Journal of Colloid and Interface Science

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Toward the Accurate Read‐out of Quantum Dot Barcodes: Design of Deconvolution Algorithms and Assessment of Fluorescence Signals in Buffer

Cited by 70 publications

References 22 publications

Exploring Primary Liver Macrophages for Studying Quantum Dot Interactions with Biological Systems

Exploring Primary Liver Macrophages for Studying Quantum Dot Interactions with Biological Systems

Facile and Rapid One‐Step Mass Preparation of Quantum‐Dot Barcodes

Preparation of quantum dots encoded microspheres by electrospray for the detection of biomolecules

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