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The antibody microarray developed has been tested for its capacity of detecting DNA-PKcs in glial cell lines and measuring cell protein phosphorylation changes caused by camptothecin-induced DNA damage with different protein kinase inhibitors in HeLa cells.Lateral flow test strips (LFTSs) are particularly attractive for assaying targets due to their ability for fast response, simple equipment, portability, and straightforward signal readout. In this work, we describe a rapid and sensitive "off-on" LFTS for point-of-care testing of glutathione (GSH) based on fluorescent quantum dots. Under the optimal conditions, a good linear relationship from 0.1 to 15 μM with a detection limit of 25 nM was obtained, and the whole process was accomplished within 10 min. In addition, this proposed LFTS was successfully used to detect GSH in HeLa cells extract.The photoelectrochemical (PEC) biosensor, in which light is utilized to excite the photoactive species and current is employed as the detection signal, is a newly appeared yet dynamically developing technique for biological analysis. Based on the assay of DNA binding proteins upon visible light irradiation, a PEC biosensor is constructed for successfully probing a DNA-protein interaction.A quantum dot (QD)-based lab-on-bead system is a unique tool for multiple analysis of cancer markers in human serum samples by using a flow cytometer. In terms of specificity and sensitivity, this method is comparable with ELISA, the "gold standard" of serological in-clinic detection of single analytes. Fluorescent microspheres encoded with QDs have been used for the quantitative detection of free and total prostate-specific antigen in human serum samples. Developed multiplex assay demonstrates a clear discrimination between serum samples from control subjects and cancer patients. The proposed QD-based method is adaptable and makes it possible to develop numerous clinical tests with decreased duration and cost for early diagnosis of various diseases.The increasing applications of quantum dots (QDs) as optic tools in life science have stimulated researchers to evaluate the effects of these nanoprobes in cell viability using a variety of methods, especially colorimetric ones. One of the most applied tests is the MTT assay. In comparison to MTT, for example, the resazurin-based method has the main advantage of not evaluating the cells directly, thus eliminating false-positive results that may arise from the overlap of the absorbances of the QD with the colorimetric compound. Therefore, herein, we describe the resazurin assay as an alternative, simple, quick, sensitivity, reproducible, and nontoxic test to evaluate the in vitro cell viability after QD exposure. Moreover, this test presents an additional advantage; the cells remain viable for complementary experimental procedures, such as cell migration or adhesion.Fluorescent semiconductor nanocrystals, known as quantum dots (QDs), and magnetic nanoparticles (MNPs) are extensively studied perspective tools for optical (fluorescence) and magnetic resonance imaging techniques. The unique optical properties, high photostability, and bright luminescence of QDs make them more promising fluorophores than the classical organic dyes. Encoding polyelectrolyte microcapsules with QDs and MNPs ensures their sensitivity to both photoexcitation and magnetic field. This chapter presents the protocol for obtaining a stimulus-sensitive delivery system based on QD- and MNP-encoded polyelectrolyte microcapsules by means of layer-by-layer self-assembly. The resultant fluorescent magnetic polyelectrolyte microcapsules are 3.4-5.5 μm in size, have a hollow structure, and are brightly fluorescent to be detected with the standard imaging equipment. Polyelectrolyte microcapsule surface bears functional groups for subsequent functionalization with vector capture molecules. The polyelectrolyte microcapsules containing combination of QDs and MNPs are advanced visualization tools, since they can be sorted in a magnetic field and at the same time are suitable for fluorescent imaging what can be applied within a wide range of diagnostic and therapeutic protocols.The ability to image single molecules in living cells has been impaired by the absence of bright, photostable fluorophores. Quantum dots (QDs) offer an attractive solution to this problem due to their exceptional photostability and brightness. Here, we describe in detail a protocol to chemically deliver functionalized QDs into the cytosol of living cells, based on cell-penetrating poly(disulfide)s (CPDs). This protocol is highly efficient and delivers hundreds of QDs per cell after incubation of cells with functionalized QDs at nanomolar concentrations. We also detail a pipeline for automated detection and tracking of diffusive QDs in living cells, which may provide a useful means to study the biophysical properties of the cytosol and their dynamics. Last, we describe a protocol for conjugating streptavidin fusion proteins to QDs, in order to permit the codelivery of QDs with functional proteins of interest into cells. The protocol has been successfully applied to a broad range of different cell types, thus offering a flexible and generalizable means to image single molecules in living cells.Single-molecule imaging has illuminated dynamics and kinetics of neuronal proteins in their native membranes helping us understand their effective roles in the brain. Here, we describe how nanometer-sized fluorescent semiconductors called quantum dots (QD) can be used to label neuronal proteins in a single QD imaging format. We detail two generalizable protocols accompanied by experimental considerations giving the user options in approach tailored to the materials and equipment available. These protocols can be modified for experiments to verify target specificity, as well as single molecule analysis such as single particle tracking and protein clustering.Brightly luminescent semiconductor quantum dots (QDs) are ideal materials for cellular imaging and analysis because of their advantageous optical properties and surface area that supports multivalent conjugation of biomolecules. An important design consideration for effective use of these materials is a hydrophilic, biocompatible surface chemistry that provides colloidal stability and minimizes nonspecific interactions with biological molecules and systems. Dextran coatings are able to satisfy these criteria. Gliocidin Despite frequent use of dextran coatings with other nanomaterials (e.g., iron oxide nanoparticles), there has been little development and application of dextran coatings for QDs. In this chapter, we describe methods for the synthesis and characterization of a dextran ligand for QDs, including preparation of an immunoconjugate via tetrameric antibody complexes (TAC). The utility of these immunoconjugates is demonstrated through immunofluorescent labeling and imaging of overexpressed human epidermal growth factor receptor 2 (HER2) on the surface of SK-BR3 breast cancer cells.
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