Quantitative measurement of multifunctional quantum dot binding to cellular targets using flow cytometry

Smith, R. A.; Giorgio, Todd D.

doi:10.1002/cyto.a.20677

Cited by 22 publications

(12 citation statements)

References 42 publications

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“…They can be injected into cells to specially deliver chemicals or stain specific parts of the cells. Both gold nanoparticles and Q-dots have been proposed to help view organelles within the cells (43,49). The unique properties of these small particles increase the signal-to-noise ratio of the staining reaction (42,43,(49)(50)(51)(52).…”

Section: Discussionmentioning

confidence: 99%

Detection of TiO₂ nanoparticles in cells by flow cytometry

et al. 2010

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Evaluation of the potential hazard of man-made nanomaterials has been hampered by a limited ability to observe and measure nanoparticles in cells. In this study, different concentrations of TiO 2 nanoparticles were suspended in cell culture medium. The suspension was then sonicated and characterized by dynamic light scattering and microscopy. Cultured human-derived retinal pigment epithelial cells (ARPE-19) were incubated with TiO 2 nanoparticles at 0, 0.1, 0.3, 1, 3, 10, and 30 lg/ml for 24 hours. Cellular reactions to nanoparticles were evaluated using flow cytometry and dark field microscopy. A FACSCalibur TM flow cytometer was used to measure changes in light scatter after nanoparticle incubation. Both the side scatter and forward scatter changed substantially in response to the TiO 2 . From 0.1 to 30 lg/ml TiO 2 , the side scatter increased sequentially while the forward scatter decreased, presumably due to substantial light reflection by the TiO 2 particles. Based on the parameters of morphology and the calcein-AM/propidium iodide viability assay, TiO 2 concentrations below 30 lg/ml TiO 2 caused minimal cytotoxicity. Microscopic analysis was done on the same cells using an E-800 Nikon microscope containing a xenon light source and special dark field objectives. At the lowest concentrations of TiO 2 (0.1-0.3 lg/ml), the flow cytometer could detect as few as 5-10 nanoparticles per cell due to intense light scattering by TiO 2 . Rings of concentrated nanoparticles were observed around the nuclei in the vicinity of the endoplasmic reticulum at higher concentrations. These data suggest that the uptake of nanoparticles within cells can be monitored with flow cytometry and confirmed by dark field microscopy. This approach may help fulfill a critical need for the scientific community to assess the relationship between nanoparticle dose and cellular toxicity Such experiments could potentially be performed more quickly and easily using the flow cytometer to measure both nanoparticle uptake and cellular health. Published

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Section: Discussionmentioning

confidence: 99%

Detection of TiO₂ nanoparticles in cells by flow cytometry

et al. 2010

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“…However, in contrast to fluorescence microscopy, flow cytometry enables quantitation of NP fluorescence but, unfortunately, not the NP mass or number. In order to make method more quantitative, toolsets that include the calibration of flow cytometer using prior measurement of the fluorescence with a known concentration of NPs have been introduced . However, these methods are currently not commonly utilized and need further development.…”

Section: Methods For Qualitative and Quantitative Analysis Of Cell‐asmentioning

confidence: 99%

Methodologies and approaches for the analysis of cell–nanoparticle interactions

Ivask

Mitchell

Malysheva

et al. 2017

WIREs Nanomed Nanobiotechnol

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How to study nanoparticle-cell interactions is the key question that puzzles researchers in the fields of nanomedicine as well as in nanotoxicology. In nanotoxicology, the amount of nanoparticles internalized by the cells or bound to the external surfaces of cells determines the toxic profile of those particles. In medical applications, cellular uptake and binding of medically effective nanoparticles decides their efficacy. Despite the importance of understanding the extent and mode of nanoparticle-cell interactions, these processes are underinvestigated, mainly due to the lack of suitable user-friendly methodologies. Here we discuss the advantages and limitations of currently available (and most advanced) microscopic, spectroscopic, and other bioanalytical methods that could be used to assess cell-nanoparticle interactions either qualitatively or quantitatively. Special emphasis is given to the methods that enable analysis and identification of nanoparticles at single-cell level, and allow intracellular localization and speciation analysis of nanoparticles. This article is categorized under: Nanotechnology Approaches to Biology > Cells at the Nanoscale Toxicology and Regulatory Issues in Nanomedicine > Toxicology of Nanomaterials.

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“…Additionally, binding of these aptamers to cell surface proteins is proposed to interfere with the function of the protein and may result in different phenotypes and potentially negatively influence the parasite. Moreover, together with trafficking analysis of GFP-labeled P. falciparum chimer proteins, aptamers identified binding to distinct parasite proteins on infected erythrocytes can be derivatized by quantum dots and used for multiparameter cytomics (73,74).…”

Section: Employing Aptamers On Parasite Infectionmentioning

confidence: 99%

Disease‐specific biomarker discovery by aptamers

Ulrich

Wrenger

2009

Cytometry Pt A

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RNA and DNA aptamers developed by an in vitro selection process, Systematic Evolution of Ligands by EXponential enrichment (SELEX), comprise a novel class of highaffinity and specific capture agents, which can be easily modified for cytometry and in vivo applications. A novel application of this technique (Cell SELEX) explores the expression of cell surface epitopes that differ between two given cell types or between healthy and diseased cells. Using whole cells as targets, aptamer libraries can be identified that bind to biomarkers expressed by target cells and not by any other cells. Aptamers have been developed that specifically interact with cell surface epitopes of trypanosomes or distinguish between the differences in molecular signature of somatic and cancer cells. Aside from its use for target cell identification by image and flow cytometry and laser-scanning microscopy, aptamers can be used for ligand-mediated purification and identification of their binding proteins in target cell membranes. In this review, we discuss an approach for the development of aptamers targeting parasitederived surface proteins of Trypanosoma and Plasmodium. ' 2009 International Society for Advancement of CytometryKey terms aptamers; Plasmodium falciparum; deconvolution SELEX; cell and target protein identification; fluorescence-labeled aptamers for clinical and cytomics application THE SELEX TECHNOLOGYThe SELEX technique (Systematic Evolution of Ligands by EXponential enrichment) initially developed by the laboratories of Larry Gold (1) and Jack Szostak (2) uses iterative in vitro selection of combinatorial RNA or DNA pools against a target molecule for the identification of high-affinity oligonucleotide ligands, known as aptamers. This methodology has been extensively used to isolate aptamers for a wide variety of peptides and proteins of therapeutic importance, including growth and coagulation factors (3). This novel class of oligonucleotide-based ligands recognizes their targets with binding specificities and affinities comparable to those of monoclonal antibodies. Moreover, aptamers are able to distinguish between protein isoforms and different conformational forms of the same protein (4,5). In most cases, aptamer binding to its target protein inhibits its biological activity, either resulting from aptamer interference with the catalytic site of an enzyme or with sites involved in ligandreceptor recognition. However, it is also possible that aptamer-target protein binding induces allosteric effects, such as changes in conformational states resulting in loss of biological activity of the target protein.Aptamers can be identified by in vitro selection against almost any target, including toxins and antigens which do not induce immune responses in host animals for antibody production. As a result, this novel class of ligands is highly promising for the development of therapeutics and biotechnological tools. The potential utility of aptamers for in vivo applications and as therapeutic agents is considerably enhanced ...

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Quantitative measurement of multifunctional quantum dot binding to cellular targets using flow cytometry

Cited by 22 publications

References 42 publications

Detection of TiO₂ nanoparticles in cells by flow cytometry

Detection of TiO₂ nanoparticles in cells by flow cytometry

Methodologies and approaches for the analysis of cell–nanoparticle interactions

Disease‐specific biomarker discovery by aptamers

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Quantitative measurement of multifunctional quantum dot binding to cellular targets using flow cytometry

Cited by 22 publications

References 42 publications

Detection of TiO2 nanoparticles in cells by flow cytometry

Detection of TiO2 nanoparticles in cells by flow cytometry

Methodologies and approaches for the analysis of cell–nanoparticle interactions

Disease‐specific biomarker discovery by aptamers

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Detection of TiO₂ nanoparticles in cells by flow cytometry

Detection of TiO₂ nanoparticles in cells by flow cytometry