Quantum Dots-based Reverse Phase Protein Microarray Page: 4 of 22
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pattern (>50 nm) poses certain problems for multiplex detections when the signals are low,
because filtering out the cross-talks between channels comes at the expense of the signal
intensity. Therefore, there is an urgent need to develop detection techniques that do not rely on
organic dyes.
Recently, reverse phase protein lysate microarrays have been reported, in which cell lysate
proteins are immobilized on nitrocellulose-coated substrate, while a dilution series of the lysates
with a dynamic range of at least one thousand-fold is used for quantification of protein [12]. This
method assesses only one protein per microarray, but it nonetheless has a great advantage
because multiple samples can be analyzed and compared side by side in a single array [13,14].
Proteins on a microarray are detected with a highly effective signal amplification procedure,
involving horseradish peroxidase (HRP)-catalyzed diaminobenzidine (DAB) chromogenesis.
DAB is a commonly used chromogen with HRP. This amplification system is based on catalyzed
reporter deposition of biotinylated tyramide [15-18]. The combination of DAB and tyramide
signal amplification (TSA) results in a brown precipitate with excellent signal-to-noise ratio [12].
Using the HRP-DAB platform, detection of proteins in single cell can be accomplished routinely
[13]. However, the HRP-DAB system is sensitive to various factors, such as temperature,
reagent quality, and specific activity of the HRP enzyme.
We investigate here the use of fluorescent semiconductor quantum dots (Qdots) as an alternative
visualization label for protein microarray studies[19]. Qdots are crystalline materials made of a
CdSe core of a few nanometers, and surrounded by a thin shell of ZnS[20]. This CdSe/ZnS
core/shell nanostructure has the ability to emit light upon UV excitation. The emission is narrow
(-20-25 nm fwhm) and can be tuned by adjusting the size of the CdSe core, due to the quantum
confinement effect. The colloidal chemistry is so well developed that it allows the synthesis of
five to six spectrally distinct colors of emission across the visible spectrum[10]. Recent progress
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Shingyoji, Masato; Gerion, Daniele; Pinkel, Dan; Gray, Joe W. & Chen, Fanqing. Quantum Dots-based Reverse Phase Protein Microarray, article, July 15, 2005; Berkeley, California. (https://digital.library.unt.edu/ark:/67531/metadc792166/m1/4/: accessed April 23, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; crediting UNT Libraries Government Documents Department.