This PDF file contains the front matter associated with SPIE Proceedings Volume 7189, including the Title Page, Copyright information, Table of Contents, Introduction (if any), and the Conference Committee listing

Nano- and cytotoxicity becomes increasingly more important with an increasing number of potential bio-medical applications for semiconductor Quantum Dots (QDs). Therefore, the frequently used CdSe-based QDs are unsuitable per-se, since cadmium is a highly toxic heavy metal and may leach out of QDs. Cadmium-free QDs have not been available for a long time, because the synthesis of e.g. monodisperes and highly crystalline InP QDs caused many problems. We report on the synthesis of InP/ZnS QDs with optical properties similar to those displayed by typical CdSe/ZnS QDs. A major break-through has been reached by addition of zinc ions into the reaction mixture. Furthermore, the transfer of the InP/ZnS QDs to water and their exploitation for bioanalytical applications are reported. It is shown that InP/ZnS QDs can be used to replace CdSe-based ones for almost any bio-medical application.

Fluorescence spectroscopy studies have shown that, at a single molecule level, fluorophore emission intensity fluctuates between bright and dark states. These fluctuations, known as blinking, limit the use of fluorophores in single molecule experiments. Statistical analysis of these intensity fluctuations has demonstrated that the dark states duration exhibits a universal heavy-tailed power law distribution in organic as well as inorganic fluorophores. However, the precise reasons underlying the blinking of single fluorophores are still matter of debate and whether it can be suppressed is not clear. Here we have synthesized CdSe/CdS core/shell quantum dots (QDs) with thick crystalline shells, which do not blink at low frame rate and established a direct correlation between shell thickness and blinking occurrences. Single fluorophore blinking and blinking statistics are thus not as universal as thought so far. We anticipate our results to help better understand the physicochemistry of single fluorophore emission and rationalize the design of other fluorophores that do not blink. The materials presented here should readily find interesting applications in biology for single molecule tracking and in photonics as robust and continuous photon emitter.

We reported for the first time that key nanocrystal quantum dot (NQD) optical properties - quantum yield, photobleaching and blinking - can be rendered independent of NQD surface chemistry and environment by growth of a very thick, defect-free inorganic shell (Chen, et al. J. Am. Chem. Soc. 2008). Here, we show the precise shell-thickness dependence of these effects. We demonstrate that 'giant-shell' NQDs can be largely non-blinking for observation times as long as 54 minutes and that on-time fractions are independent of experimental time-resolution from 1-200 ms. These effects are primarily demonstrated on (CdSe)CdS (core)shell NQDs, but we also show that alloyed shells comprising Cd_xZn_1-xS and terminated with a non-cytotoxic ZnS layer exhibit similar properties. The mechanism for suppressed blinking and dramatically enhanced stability is attributed to both effective isolation of the NQD core excitonic wavefunction from the NQD surface, as well as a quasi-Type II electronic structure. The unusual electronic structure provides for effective spatial separation of the electron and hole into the shell and core, respectively, and, thereby, for reduced efficiencies in non-radiative Auger recombination.

The response of solubilized quantum dot solutions to visible or UV irradiation is highly variable, and contradictory reports exist in the literature. Using several different preparations of core CdSe, core-shell CdSe/ZnS, and CdTe quantum dots (QDs), we investigated the time-resolved photoluminescence as a function of 400 nm irradiation. We found that photoenhancement and photodegradation were highly dependent upon irradiation power, with the QDs being highly stable at fluences of < 2 mW. However, great variability was seen among independent preparations of QDs, with fresher dots showing greater photostability than those that had been aged in organic solvent. Conjugation of dopamine to the QDs also led to variable effects, with some batches showing lifetime enhancement upon conjugation and others suppression. In all cases, QD-dopamine conjugates showed increased lifetimes upon irradiation, up to a maximum effect at ~ 5 min post irradiation at 2.4 mW. The antioxidant beta-mercaptoethanol also affected different batches of QDs differently; it prevented photoenhancement with certain batches but not others. We propose a mechanism of photoenhancement and surface oxidation that relates the variability to the number of solubilising groups on the QD surface. The potential of photoenhancement as a sensing mechanism in cells is proposed.

Quantum dots are nanometre-sized semiconductor particles exhibiting unique size-dependent electronic properties. In order to passivate the nanocrystals surface and to protect them from oxidation, we grow a shell composed of a second semiconductor with a larger bandgap on the core (for example a core / shell CdS / ZnS). However, the lattice mismatch between the two materials (typically 7% between ZnS and CdS) induces mechanical stress which can lead to dislocations. To better understand these mechanisms, it is important to be able to measure the pressure induced on the semiconductor core. We used a nanocrystal doped with manganese ions Mn²⁺, which provide a phosphorescence signal depending on the local pressure. A few dopant atoms per nanoparticle were placed at controlled radial positions in a ZnS shell formed layer by layer. The experimental pressure measurements are in very good agreement with a simple spherically symmetric elastic continuum model[1]. Using manganese as a pressure gauge could be used to better understand some structural phenomena observed in these nanocrystals, such as crystalline phases transition, or shell cracking.

Colloidal synthesis of cerium-doped lanthanum fluoride nanocrystals is reported. The nanocrystals were characterized by transmission electron microscopy, energy-dispersive X-ray spectroscopy, steady-state UV-VIS optical absorption and photoluminescence (PL) spectroscopy, and by PL lifetime measurements. Cerium doping concentration was optimized for the maximum PL intensity. Radiation hardness of the synthesized nanocrystals was tested using a ¹³⁷Cs 662-keV gamma source. Finally, scintillation was observed from the cerium-doped lanthanum fluoride nanocrystals exposed to low-level gamma radiation.

Fluorescence is increasingly used for in vivo imaging and has provided remarkable results. Howerver this technique presents several limitations, especially due to tissue autofluorescence under external illumination and weak tissue penetration of low wavelength excitation light. We have developed an alternative optical imaging technique using persistent luminescent nanoparticles suitable for small animal imaging. These nanoparticles can be excited before the injection, and their in vivo distribution can be followed in real-time for several hours. Chemical modifications of their surface is possible leading to lung or liver targeting, or to long-lasting blood circulation.

The use of semiconductor luminescent quantum dots for the labeling of biomolecules is rapidly expanding, however it still requires facile methods to attach functional globular proteins to biologically optimized quantum dots. Here we discuss the development of controlled variable length peptidyl linkers to attach biomolecules to poly(ethylene) glycol (PEG) coated quantum dots for both in vitro and in vivo applications. The peptides chosen, β-sheets and alpha helices are appended to polyhistidine sequences and this allows for control of the ratio of peptide bioconjugated to QD and the distance from QD to the biomolecule. Recombinant DNA engineering, bacterial peptide expression and Ni-NTA purification of histidine labeled peptides are utilized to create the linkers. Peptide length is confirmed by in vitro fluorescent resonance energy transfer (FRET).

One of the most prominent research areas in nanotechnology is the development of nanoparticle systems for biomedical applications. This is founded upon the expectation that such species could ultimately be imbued with multiple simultaneous functions, such as the presentation of a therapeutic payload or diagnostic sensor for in vivo trafficking to desired cell types. In recent years, semiconductor quantum dots (QDs) have been actively explored as novel display systems, because of their unique photophysical properties. Using an aniline-mediated hydrazone coupling, a polyhisitidine-appended peptide was derivatized with a DNA strand and successfully self-assembled to QDs, yielding nanoparticles displaying up to approximately 15 peptide/DNA conjugates. This ligation method is a viable chemistry for displaying biomolecules, because of the orthogonality of the ketone and hydrazine moieties to most biological functionality and the reaction can be performed under mild conditions in aqueous media. The modified QDs were further characterized by gel electrophoresis, and microarray studies; showing the self-assembly was successful and the DNA strands were still available for hybridization with a complement sequence.

Homogeneous immunoassays have the benefit that they do not require any time-consuming separation steps. FRET is one of the most sensitive homogeneous methods used for immunoassays. Due to their extremely strong absorption over a broad wavelength range the use of quantum dots as FRET acceptors allows for large Foerster radii, an important advantage for assays in the 5 to 10 nm distance range. Moreover, because of their size-tunable emission, quantum dots of different sizes can be used with a single donor for the detection of different analytes (multiplexing). As the use of organic dyes with short fluorescence decay times as donors is known to be inefficient with quantum dot acceptors, lanthanide complexes with long luminescence decays are very efficient alternatives. In this contribution we present the application of commercially available biocompatible CdSe/ZnS core/shell quantum dots as multiplexing FRET acceptors together with a single terbium complex as donor in a homogeneous immunoassay system. Foerster radii of 10 nm and FRET efficiencies of 75 % are demonstrated. The high sensitivity of the terbium-toquantum dot FRET assay is shown by sub-100-femtomolar detection limits for two different quantum dots (emitting at 605 and 655 nm) within the same biotin-streptavidin assay. Direct comparison to the FRET immunoassay "gold standard" (FRET from Eu-TBP to APC) yields a three orders of magnitude sensitivity improvement, demonstrating the big advantages of quantum dots not only for multiplexing but also for highly sensitive nanoscale analysis.

Luminescent colloidal semiconductor quantum dots (QDs) have unique optical and photonic properties and are highly sensitive to charge transfer in their surrounding environment. In this study we used synthetic peptides as physical bridges between CdSe-ZnS core-shell QDs and some of the most common redox-active metal complexes to understand the charge transfer interactions between the metal complexes and QDs. We found that QD emission underwent quenching that was highly dependent on the choice of metal complex used. We also found that quenching traces the valence or number of metal complexes brought into close proximity of the nanocrystal surface. Monitoring of the QD absorption bleaching in the presence of the metal complex provided insight into the charge transfer mechanism. The data suggest that two distinct charge transfer mechanisms can take place. One directly to the QD core states for neutral capping ligands and a second to surface states for negatively charged capping ligands. A basic understanding of the proximity driven charge-transfer and quenching interactions allowed us to construct proteolytic enzyme sensing assemblies with the QD-peptide-metal complex conjugates.

Quantum dots (QDs) are unique probes due to their special properties (brightness, photostability, narrowband emission and broadband absorption), and excellent bio(chemical)compatibility for imaging structures and functions of living cells. When functionalized with ligands, they enable the recognition of specific targets and the tracking of dynamic processes for extended periods of time, detecting biomolecules with a sensitivity extending to the single molecule level. Thus, devices and probes based on such nanoparticles are very powerful tools for studying essential processes underlying the functions and regulation of living cells. Here we present nanosensors and nanoactuators based on QDs in which the multivalency of these particles plays an essential role in the functionality and sensing characteristics of the nanodevices. Two examples are discussed, the first being pH nanosensors based on the interplay of the multivalency and energy transfer between the nanoparticles and small molecules on their surface, and the second nanoactuators in which a controlled number of molecules of the amyloid protein α-synuclein (AS) specifically regulate the aggregation of fluorescently labeled bulk AS protein both in vitro and in live cells.

Individual microspheres labeled with a unique barcode and a surface-bound probe are able to provide multiplexed biological assays in a convenient and high-throughput format. Typically, barcodes are created by impregnating microspheres with several colors of fluorophores mixed at different intensity levels. The number of barcodes is limited to hundreds primarily due to variability in fluorophore loading and difficulties in compensating for signal crosstalk. We constructed a molecular barcode based on differences in lifetimes rather than intensities. Lifetime-based measurements have an advantage in that signal from neighboring channels is reduced (because signal intensities are equal) and may be mathematically deconvoluted. The excited state lifetime of quantum dots (QDs) was systematically altered by attaching a variable number of quencher molecules to the surface. We have synthesized a series of ten QDs with distinguishable lifetimes all emitting at the same wavelength. The QDs were loaded into microspheres to determine the expected signal intensities. The uncertainty in lifetimes as a function of the interrogation time was determined. An acceptable standard deviation (3%) was obtained with a measurement time of approximately 10-30 μsec. Currently, we are expanding these studies to include multiple wavelengths and determining the maximal number of barcodes for a given spectral window.

Can quantum dots (QDs) act as relevant intracellular probes to investigate routing of ligands in live cells? To answer this question we studied intracellular trafficking of QDs that were coupled to the plant toxin ricin, Shiga toxin or the ligand transferrin (Tf) by confocal fluorescence microscopy in three different cell lines. The Tf:QDs were internalized but instead of being recycled they accumulated within endosomes in all cell lines. However, for the HEp-2 and SW480 cells a higher fraction colocalized with a lysosomal marker as compared with HeLa cells. The Shiga:QD bioconjugate was internalized slowly and with poor efficiency in the HEp-2 and SW480 cells as compared with HeLa cells, and was not routed to the Golgi apparatus in any of the cell lines. The internalized ricin:QD bioconjugates localized to the same endosomes as ricin itself, but could in contrast to ricin not be visualized in the Golgi apparatus. Importantly, we find that the endosomal accumulation of either ricin:QDs or transferrin:QDs affects endosome-to-Golgi transport of both ricin and Shiga toxin: Transport of ricin was reduced whereas transport of Shiga toxin was increased. In conclusion, the data from different cells reveal that in general these ligand-coupled QD nanoparticles are arrested within endosomes, and somehow perturb the normal endosomal sorting in cells.

During the last decade, there has been great deal of interest in the self-assembly fabrication of hybrid materials from inorganic nanoparticles and biomolecules. Nanoparticles are similar in size range to many common biomolecules, thus, nanoparticles appear to be natural companions in hybrid systems. At present, it is straightforward to control and modify properties of nanostructures to better suit their integration with biological systems; for example, controlling their size, modifying their surface layer for enhanced aqueous solubility, biocompatibility, or biorecognition. A particularly desirable target for therapeutic uses is the cell nucleus, because the genetic information is there. We review in this article the synthesis developed by our research group of water-soluble gold nanoparticles and CdS nanocrystals functionalized with a Tat protein-derived peptide sequence by straightforward and economical methodologies. The particles were subsequently tested in vitro with a human fibroblast cell line using optical and transmission electron microscopy to determine the biocompatibility of these nanoparticles and whether the functionalization with the cell penetrating peptide allowed particles to transfer across the cell membrane and locate into the nucleus.

Gene therapy is an attractive approach to supplement a deficient gene function. Although there has been some success with specific gene delivery using various methods including viral vectors and liposomes, most of these methods have a limited efficiency or also carry a risk for oncogenesis. Fluorescent nanoparticles, such as nanocrystal quantum dots (QDs), have potential to be applied to molecular biology and bioimaging, since some nanocrystals emit higher and longer lasting fluorescence than conventional organic probes do. We herein report that quantum dots (QDs) conjugated with nuclear localizing signal peptides (NLSP) successfully introduced the gene-fragments with promoter elements, which promoted the expression of the enhanced green fluorescent protein (eGFP) gene in mammalian cells. The expression of eGFP protein was observed when the QD/geneconstruct was added to the culture media. The gene-expression efficiency varied depending on multiple factors around QDs, such as 1) the reading direction of gene fragments, 2) the quantity of gene fragments attached on the surface of QD-constructs, 3) the surface electronic charges varied according to the structure of QD/gene-constructs, and 4) the particle size of QD/gene complex varied according to the structure and amounts of gene fragments. Using this QD/geneconstruct system, eGFP protein could be detected 28 days after the gene-introduction whereas the fluorescence of QDs was disappeared. This system therefore provides another method for the intracellular delivery of gene-fragments without using either viral vectors or specific liposomes. These results suggest that inappropriate treatment and disposal of QDs may still have risks to the environmental pollution including human health under certain conditions. Here we propose the further research for the immune and physiological responses in not only immune cells but also other cells, in order to clear the effect of all other nanoscale products as well as nanocrystal QDs.

Luminescent semiconductor quantum dots (QDs) possess several unique optical properties that suggest they will be superior reagents compared to traditional organic fluorophores for applications such as the labeling of subcellular structures and the sensing of biological processes within living cells. Chief among these properties are their 1) high quantum yields, 2) broad absorption spectra coupled with narrow symmetric, size-tunable emissions and 3) the ability to excite multiple QD populations at a single wavelength removed from emission. This latter attribute presents the exciting possibility of multiplexed or "multicolor" intracellular imaging and sensing. In order for QDs to reach their full potential as intracellular imaging and sensing reagents, however, facile and robust methodologies for delivering QDs in a controlled manner to specific subcellular locations must be developed. We have investigated a number of strategies to achieve the intracellular delivery of QDs including peptide-mediated, polymer-mediated, and microinjection based methods. In particular, the ability to selectively deliver biofunctionalized QDs to specific intracellular compartments is being targeted. Additionally, long-term QD intracellular QD fate, stability and toxicity are also concomitantly being examined. Cellular delivery experiments utilizing these various schemes will be highlighted and the relative advantages and disadvantages of each approach will be discussed.

Probing the spatiotemporal response of individual intracellular proteins and multi-peptide complexes is essential in understanding the integrated response of cells. Although dynamic information can be captured using optical microscopy, most conventional spatial resolutions are limited to around 200 nm, which is significantly greater than the size of molecules. One mode of microscopy that overcomes this resolution limitation is the electron microscope, which enables in situ protein labeling and allows for single or sub-nanometer resolution to be obtained. Transmission electron microscopy though is limited by the inability to capture dynamic molecular responses. Here, we have demonstrated the ability to use quantum dots for both modes of microscopy through a single labeling technology, which allows both dynamic and high resolution visualization with optical and electron microscopy. We visualized core-shell CdSe/ZnS quantum dots within Dictyostelium discoideum using both microscopy modes through a bacterial nutrient protocol, which enables the quantum dots to enter living cells without the need of an artificial transporter system for assisted internalization. Optical imaging was first used to visualize the spatiotemporal behavior of actin filaments using phalloidin conjugated quantum dots. The same cells were then imaged using a transmission electron microscope to examine the detailed intracellular distribution down to a single nanometer size scale. These results have potential applications in a variety of areas including biophysics, cell motility, cancer metastasis, and cell structure.

Single particle tracking (SPT) provides information about the microscopic motions of individual particles in live cells. We applied SPT to study the diffusion of membrane transport proteins in cell plasma membranes in which individual proteins are labeled with quantum dots at engineered extracellular epitopes. Software was created to deduce particle diffusive modes from quantum dot trajectories. SPT of aquaporin (AQP) water channels and cystic fibrosis transmembrane conductance regulator (CFTR) chloride channels revealed several types of diffusion. AQP1 was freely mobile in cell membranes, showing rapid, Brownian-type diffusion. The full-length (M1) isoform of AQP4 also diffused rapidly, though the diffusion of a shorter (M23) isoform of AQP4 was highly restricted due to its supermolecular assembly in raft-like orthogonal arrays. CFTR mobility was also highly restricted, in a spring-like potential, due to its tethering to the actin cytoskeleton through PDZ-domain C-terminus interactions. The biological significance of regulated diffusion of membrane transport proteins is a subject of active investigation.

We demonstrate a novel multi-photon imaging modality based on the detection of four-wave mixing (FWM) from colloidal nanoparticles. Four-wave mixing is a third-order signal which can be excited and detected in resonance with the ground-state excitonic transition of CdSe/ZnS quantum dots. The coherent FWM signal is detected interferometrically to reject incoherent backgrounds for improved image contrast compared to fluorescence methods. We measure transversal and axial resolutions of 140nm and 590nm respectively, significantly beating the one-photon diffraction limit. We also demonstrate optical imaging of quantum-dot-labeled Golgi structures of HepG2 cells.

Quantum dots (Qdots) have become ubiquitous in biomedical research due to their excellent brightness, photostability, monodispersity, and fluorescent yield. Furthermore, they have become increasingly useful as imaging agents which are valuable for answering molecular questions in living subjects. However, little is currently known about how nanoparticles such as Qdots interact at the microscale within the vasculature and tumor microenvironments in living subjects. In order to further our understanding of the dynamic processes involved in Qdot targeting in the intact tumor, we developed an in vivo binding assay to visualize and fully elucidate this approach using a variety of animal models and tumor types. We employed argine-glycine-aspartic acid (RGD) peptides to specifically target the α_vβ₃ integrins which are expressed on the surface of endothelial cells comprising newly formed or forming blood vessels; RGD peptides were conjugated to the Qdot surface. Exploiting intravital microscopy with subcellular-level resolution, we directly observed and recorded the binding of nanoparticle conjugates in two different murine models, using three different tumor cell lines. Using this generalizable approach, we learned that RGD-qdots unexpectedly bind to tumor blood vessels in all models tested only as aggregates rather than individually. Understanding these issues on the microscale using such techniques will provide a platform for the rational design of molecularly-targeted nanoparticles including Qdots. This is critical for nanoparticles to become a valuable research tool with the potential to become clinically valuable imaging and therapeutic agents, particularly for ensuring regulatory approval of such nanoparticles.

Nanoparticle toxicity is of interest for the design of anti-bacterial and anti-cancer agents, and also for development of regulations for their safe handling and disposal. However, the same reactivity that makes nanoparticles show unique optical properties also makes them interact with standard cytotoxicity agents, causing false positive and/or false negative results. We discuss which cytotoxicity assays are most likely to work with nanoparticles and which are to be avoided, and present some results on comparative cytotoxicity of different types of conjugates.

Cerium-doped lanthanum fluoride colloidal nanocrystals offer a way to improve external radiation therapy through the enhanced absorption of high energy photons, as well as through the emission of UV light in the presence of radiation, providing a second cell killing mechanism. Lanthanum fluoride nanocrystals doped with 10% cerium were anhydrously synthesized in methanol as platelets 10-12 nm in diameter and 4-6 nm thick. The nanocrystals were characterized by transmission electron microscopy, energy dispersive spectroscopy (EDS), and by steady state UV-visible optical absorption and photoluminescence spectroscopy. Using an incoming gamma flux from a ¹³⁷Cs source and a Fricke dosimeter solution to measure absorbed energy, a 55% enhancement of absorbed dose was measured for a 1.2 mg/ml loading of nanocrystals over exposure range from one to four kiloroentgens.

The development of microbiological biofilms greatly reduces the efficacy of antibiotic therapies and is a serious problem in chronic infection and for implantable medical devices. We investigated the potential of superparamagnetic nanoparticles to increase transport through in vitro models of alginate biofilms. An in vitro alginate biofilm model was developed to mimic the composition of in vivo samples of P. aeruginosa infections. Transport through this model biofilm was performed using both bulk diffusion methods and single particle tracking techniques in the presence and absence of an external magnetic field. Bulk diffusion of nanoparticles through the biofilm was significantly enhanced in the presence of a magnetic field, both visually and quantitatively. Nanoparticle trajectories also showed transport increases were significantly higher when magnetic fields were applied. We also showed that surface chemistry (cationic, anioni, or neutral) of the nanoparticles significantly influenced transport rates. Finally, nanoparticle size also influenced the transport rates and variability of transport rates through the biofilm. In these first studies using magnetic nanoparticles in bacterial biofilms, we demonstrate that transport enhancement can be achieved and further studies are warranted.

The growing presence of quantum dots (QD) in a variety of biological, medical, and electronics applications means an increased risk of human exposure in manufacturing, research, and consumer use. However, very few studies have investigated the susceptibility of skin to penetration of QD - the most common exposure route- and the results of those that exist are conflicting. This suggests that a technique allowing determination of skin barrier status and prediction of skin permeability to QD would be of crucial interest as recent findings have provided evidence of in vitro cytotoxicity and long-term in vivo retention in the body for most QD surface chemistries. Our research focuses on barrier status of the skin (intact and with ultraviolet radiation induced barrier defect) and its impact on QD skin penetration. These model studies are particularly relevant to the common application condition of NP containing sunscreen and SPF cosmetics to UV exposed skin. Herein we present our initial efforts to develop an in vivo model of nanoparticle skin penetration using the SKH-1 hairless mouse with transepidermal water loss (TEWL) to evaluate skin barrier status and determine its ability to predict QD penetration. Our results show that ultraviolet radiation increases both TEWL and skin penetration of QD. Additionally, we demonstrate cytotoxic potential of QD to skin cells using a metastatic melanoma cell line. Our research suggests future work in specific targeting of nanoparticles, to prevent or enhance penetration. This knowledge will be used to develop powerful therapeutic agents, decreased penetration cosmetic nanoparticles, and precise skin cancer imaging modalities.

Carbon group quantum dots (QDs) such as carbon, silicon and germanium, have potential for biomedical applications such as bio-imaging markers and drug delivery systems and are expected to demonstrate several advantages over conventional fluorescent QDs such as CdSe, especially in biocompatibility. We assessed biocompatibility of newly manufactured silicon QDs (Si-QDs), by means of both MTT assay and LDH assay for HeLa cells in culture and thereby detected the cellular toxicity by administration of high concentration of Si-QD (>1000 μg/mL), while we detected the high toxicity by administration of over 100 μg/mL of CdSe-QDs. As a hypothesis for the cause of the cellular toxicity, we measured oxy-radical generation from the QDs by means of luminol reaction method. We detected generation of oxy-radicals from the Si-QDs and those were decreased by radical scavenger such as superoxide dismutase (SOD) and N-acetyl cysteine (NAC). We concluded that the Si-QD application to cultured cells in high concentration led cell membrane damage by oxy-radicals and combination usage with radical scavenger is one of the answers.