Synthesis and characterisation of quantum dots coupled to mycolic acids as a water-soluble fluorescent probe for potential lateral flow detection of antibodies and diagnosis of tuberculosis

This work explores the potential use of cadmium-based quantum dots (QDs) coupled to mycolic acids (MAs) as a fluorescent probe to detect anti-MA antibodies which are biomarkers for tuberculosis (TB). The use of free MAs as antigens for the serodiagnosis of TB is known but has not been developed into a point of care test. This study focuses on the synthesis, solubility, and lateral flow of QDs coupled to MAs. Water-soluble CdSe/ZnS QDs capped with l -cysteine were synthesised and covalently coupled to MAs via amide linkages to form a water-soluble fluorescent probe: MA-CdSe/ZnS QDs. The MA-CdSe/ZnS QDs showed broad absorption bands and coupling, confirmed by the presence of amide bonds in the Fourier-transform infrared (FTIR) spectrum, resulting in a blue shift in fluorescence. Powder X-ray diffraction (XRD) revealed a shift and increase in the number of peaks for MA-CdSe/ZnS QDs relative to the L-cys-CdSe/ZnS QDs, suggesting that coupling changed the crystal structure. The average particle size of MA-CdSe/ZnS QDs was ~3.0 nm. Visual paper-based lateral flow of MA-CdSe/ZnS QDs was achieved on strips of nitrocellulose membrane with both water and membrane blocking solution eluents. The highly fluorescent MA-CdSe/ZnS QDs showed good water solubility and lateral flow, which are important properties for fluorescence sensing applications.     

Enhancement of cell internalization and photostability of red and green emitter quantum dots upon entrapment in novel cationic nanoliposomes

Two quantum dots (QDs), a green emitter, CdSe and a red emitter, CdSe with ZnS shell are encapsulated into novel liposomes in two different formulations including cationic liposomes. Quantum dots have proven themselves as powerful inorganic fluorescent probes, especially for long‐term, multiplexed imaging and detection. Upon delivery into a cell, in endocytic vesicles such as endosomes, their fluorescence is quenched. We have investigated the potential toxic effects, photophysical properties and cell internalization of QDs in new formulation of liposomes as an in vitro vesicle model. Entrapment of QDs into liposomes is brought about with a decrease in their intrinsic fluorescence and toxicities and an increase in their photostability and lifetime. The biomimetic lipid bilayer of liposomes provides high biocompatibility, thereby enhancing the effectiveness of fluorescent nanoparticles for biological recognition in vitro and in vivo. The prepared lipodots could effectively prevent QDs from photo‐oxidation during storage and when exposed to ultraviolet (UV) light. Moreover, the flow cytometry of HEK 293 T cells showed that the cell internalization of encapsulated QDs in (DSPC/CHO/DOPE/DOAB) liposome is enhanced 10 times compared with non‐encapsulated QD (bare QDs).     

聚乙二醇介导鹅掌楸悬浮细胞与CdSe／ZnS量子点纳米颗粒共孵育的互作特征

利用纳米材料介导的药物靶向治疗和动物细胞转基因等相关研究,日益受到人们的关注．但植物因存在细胞壁的障碍,无论原位还是离体细胞培养条件下,利用纳米技术进行基因转移均存在很大难度．因此设想,如通过纳米颗粒材料物理尺寸的改变和表面化学修饰,能改变纳米颗粒与植物细胞壁界面上的生物物理或生物化学特征,从而有利于纳米颗粒材料穿越植物细胞壁进入植物细胞,将对推动纳米技术在植物转基因领域中的应用产生重要意义．根据以上设想,研究了不同的共孵育时间和温度等条件下,杂交鹅掌楸的胚性悬浮细胞与经不同表面化学修饰的CdSe／ZnS纳米颗粒之间相互作用过程的细胞生物学特征,以及CdSe／ZnS量子点的细胞毒性．结果表明,在共孵育后3h以内,激光共聚焦显微镜和电子扫描显微镜下,均可观察到经表面后修饰带正电荷的CdSe／ZnS纳米颗粒．同时,胞吞进入细胞内部的表面携带正电荷的CdSe／ZnS纳米颗粒的量明显与共培养时间、温度有明显的依赖关系,表明它们可以通过细胞的液相胞吞作用进入杂交鹅掌楸细胞内,且不影响细胞的活性;而表面带负电荷的CdSe／ZnS纳米颗粒则主要聚集在细胞外壁附近．在培养溶液中添加20％（质量比）聚乙二醇,可进一步提高鹅掌楸细胞胞吞CdSe／ZnS纳米颗粒的量和减轻CdSe／ZnS纳米颗粒的细胞毒性．本研究表明,以表面携带正电荷的CdSe／ZnS量子点纳米材料作为基因载体,在植物悬浮细胞的转基因研究和应用中具有广泛的前景．     

Quantifying engineered nanomaterial toxicity: comparison of common cytotoxicity and gene expression measurements

Background

When evaluating the toxicity of engineered nanomaterials (ENMS) it is important to use multiple bioassays based on different mechanisms of action. In this regard we evaluated the use of gene expression and common cytotoxicity measurements using as test materials, two selected nanoparticles with known differences in toxicity, 5 nm mercaptoundecanoic acid (MUA)-capped InP and CdSe quantum dots (QDs). We tested the effects of these QDs at concentrations ranging from 0.5 to 160 µg/mL on cultured normal human bronchial epithelial (NHBE) cells using four common cytotoxicity assays: the dichlorofluorescein assay for reactive oxygen species (ROS), the lactate dehydrogenase assay for membrane viability (LDH), the mitochondrial dehydrogenase assay for mitochondrial function, and the Comet assay for DNA strand breaks.

Results

The cytotoxicity assays showed similar trends when exposed to nanoparticles for 24 h at 80 µg/mL with a threefold increase in ROS with exposure to CdSe QDs compared to an insignificant change in ROS levels after exposure to InP QDs, a twofold increase in the LDH necrosis assay in NHBE cells with exposure to CdSe QDs compared to a 50% decrease for InP QDs, a 60% decrease in the mitochondrial function assay upon exposure to CdSe QDs compared to a minimal increase in the case of InP and significant DNA strand breaks after exposure to CdSe QDs compared to no significant DNA strand breaks with InP. High-throughput quantitative real-time polymerase chain reaction (qRT-PCR) data for cells exposed for 6 h at a concentration of 80 µg/mL were consistent with the cytotoxicity assays showing major differences in DNA damage, DNA repair and mitochondrial function gene regulatory responses to the CdSe and InP QDs. The BRCA2, CYP1A1, CYP1B1, CDK1, SFN and VEGFA genes were observed to be upregulated specifically from increased CdSe exposure and suggests their possible utility as biomarkers for toxicity.

Conclusions

This study can serve as a model for comparing traditional cytotoxicity assays and gene expression measurements and to determine candidate biomarkers for assessing the biocompatibility of ENMs.

     

Hydrothermal synthesis for high‐quality glutathione‐capped CdxZn1 – xSe and CdxZn1 – xSe/ZnS alloyed quantum dots and its application in Hg(II) sensing

High‐quality Cd_xZn_{1 – x}Se and Cd_xZn_{1 – x}Se/ZnS core/shell quantum dots (QDs) emitting in the violet–green spectral range have been successfully prepared using hydrothermal methods. The obtained aqueous Cd_xZn_{1 – x}Se and Cd_xZn_{1 – x}Se/ZnS QDs exhibit a tunable photoluminescence (PL) emission (from 433.5 nm to 501.2 nm) and a favorable narrow photoluminescence bandwidth [full width at half maximum (FWHM): 30–42 nm]. After coating with a ZnS shell, the quantum yield increases from 40.2% to 48.1%. These Cd_xZn_{1 – x}Se and Cd_xZn_{1 – x}Se/ZnS QDs were characterized by transmission electron microscopy, X‐ray diffraction, X‐ray photoelectron spectroscopy and Fourier transform infrared (FTIR) spectroscopy. To further understand the alloying mechanism, the growth kinetics of Cd_xZn_{1 – x}Se were investigated through measuring the fluorescence spectra and X‐ray diffraction spectra at different growth intervals. The results demonstrate that the inverted ZnSe/CdSe core/shell structure is formed initially after the injection of Cd²⁺. With further heating, the core/shell structured ZnSe/CdSe is transformed into alloyed Cd_xZn_{1 – x}Se QDs with the diffusion of Cd²⁺ into ZnSe matrices. With increasing the reaction temperature from 100 °C to 180 °C, the duration time of the alloying process decreases from 210 min to 20 min. In addition, the cytotoxicity of Cd_xZn_{1 – x}Se and Cd_xZn_{1 – x}Se/ZnS QDs were investigated. The results indicate that the as‐prepared Cd_xZn_{1 – x}Se/ZnS QDs have low cytotoxicity, which makes them a promising probe for cell imaging. Finally, the as‐prepared Cd_xZn_{1 – x}Se/ZnS QDs were utilized to ultrasensitively and selectively detect Hg²⁺ ions with a low detection limit (1.8 nM).     

Quenching of Fluorescence from CdSe/ZnS Nanocrystal QDs Near Copper Nanoparticles in Aqueous Solution

Significant quenching of fluorescence from CdSe/ZnS nanocrystal quantum dots (QDs) coated with mercaptoundecanoic ligands has been realized by copper nanoparticles (NPs). (a) Static quenching in the electrostatic association between the CdSe/ZnS QDs and cetyltrimethylammonium bromide-coated Cu NPs and (b) dynamic quenching of the same nanocrystals by polyvinylpyrrolidone-coated Cu NPs were studied. In both cases, the quenching of fluorescence from the CdSe/ZnS nanocrystals is sensitive to nanomolar concentrations of the copper NPs, and the quenching efficiency increases as spectral overlap between the CdSe/ZnS emission and the copper nanoparticle absorption increases. This suggests that the observed quenching is a result of energy transfer processes. These findings open new avenues for the utilization of Cu NPs in energy transfer-based applications.     

Preparation and Biological Effect of Nucleotide-Capped CdSe/ZnS Quantum Dots on Tetrahymena thermophila

In this paper, we described the preparation and characterization of different types of modified CdSe/ZnS quantum dots (QDs) and explored the biological effects of QDs with different surface modifications on the whole growth of unicellular protozoan Tetrahymena thermophila BF(5) using a thermal activity monitor air isothermal microcalorimeter. Our results demonstrated that adenosine 5'-monophosphate (AMP) showed stronger interaction with QDs than other types of nucleotide. AMP-QDs could stimulate the growth of T. thermophila while mercaptoacetic acid-capped CdSe/ZnS quantum dots inhibited it. In addition, the population density determination and fluorescence imaging of T. thermophila BF(5) also confirmed the results obtained from microcalorimetry. It is believed that this approach will provide a more convenient methodology for the kinetics and thermodynamics of microorganism when coexisting with QDs in real time, and all of which are very significant to understanding the effect of QDs to organism.     

Fluid Phase Endocytic Uptake of Artificial Nano-Spheres and Fluorescent Quantum Dots by Sycamore Cultured Cells: Evidence for the Distribution of Solutes to Different Intracellular Compartments

Fluid phase endocytic uptake of external solutes in plant cells was further substantiated using artificial polystyrene nano-spheres (40 nm) and CdSe/ZnS quantum dots (20 nm). Both types of artificial nano-particles were taken up by sycamore-cultured cells. However, whereas polystyrene nano-spheres were delivered to the central vacuole, CdSe/ZnS nano-dots were sequestered into cytoplasmic vesicular structures. Using dextran-Texas Red (m.w. 3,000; d-TR) as additional marker, confocal micrographs confirmed the distinct topographic distribution of CdSe/ZnS quantum dots within the cell. Initially, d-TR and CdSe/ZnS quantum dots colocalized within cytoplasmic vesicles. After 18 h incubation, d-TR was distinctly localized in the vacuole whereas CdSe/ZnS quantum dots remained sequestered in cytoplasmic membranous compartments. The data provide a first evidence for the rapid distribution of solutes taken up by endocytosis to distinct intracellular compartments.Key Words: apoplast, assimilate partitioning, endocytosis, protoplasts, sucrose transport, sucrose uptake, vacuole     

3D cellular spheroids as tools for understanding carboxylated quantum dot behavior in tumors

Background

Monolayer cell cultures have been considered the most suitable technique for in vivo cellular experiments. However, a lot of cellular functions and responses that are present in natural tissues are lost in two-dimensional cell cultures. In this context, nanoparticle accumulation data presented in literature are often not accurate enough to predict behavior of nanoparticles in vivo. Cellular spheroids show a higher degree of morphological and functional similarity to the tissues.

Exome sequencing and metabolomic analysis of a chronic kidney disease and hearing loss patient family revealed RMND1 mutation induced sphingolipid metabolism defects

Mitochondrial disorders (MIDs) shows overlapping clinical presentations owing to the genetic and metabolic defects of mitochondria. However, specific relationship between inherited mutations in nuclear encoded mitochondrial proteins and their functional impacts in terms of metabolic defects in patients is not yet well explored. Therefore, using high throughput whole exome sequencing (WES), we screened a chronic kidney disease (CKD) and sensorineural hearing loss (SNHL) patient, and her family members to ascertain the mode of inheritance of the mutation, and healthy population controls to establish its rare frequency. The impact of mutation on biophysical characteristics of the protein was further studied by mapping it in 3D structure. Furthermore, LC-MS tandem mass spectrophotometry based untargeted metabolomic profiling was done to study the fluctuations in plasma metabolites relevant to disease causative mutations and kidney damage. We identified a very rare homozygous c.631G > A (p.Val211Met) pathogenic mutation in RMND1 gene in the proband, which is inherited in an autosomal recessive fashion. This gene is involved in the mitochondrial translational pathways and contribute in mitochondrial energy metabolism. The p.Val211Met mutation is found to disturb the structural orientation (RMSD is −2.95 Å) and stability (ΔΔG is −0.552 Kcal/mol) of the RMND1 protein. Plasma metabolomics analysis revealed the aberrant accumulation of metabolites connected to lipid and amino acid metabolism pathways. Of these metabolites, pathway networking has discovered ceramide, a metabolite of sphingolipids, which plays a role in different signaling cascades including mitochondrial membrane biosynthesis, is highly elevated in this patient. This study suggests that genetic defects in RMND1 gene alters the mitochondrial energy metabolism leading to the accumulation of ceramide, and subsequently promote dysregulated apoptosis and tissue necrosis in kidneys.     

量子点在生物医学中的应用

半导体量子点是无机纳米结晶,构成于硒化镉核心和硫化锌外壳.这种荧光标记物的发射光强是常用有机荧光染料的20倍,稳定性是其100倍.量子点的发射波长取决于核心粒子的大小,而每一种单色量子点的发射波长窄而对称.这些光学特性使量子点在医学诊断、药物的高速筛选以及基因和蛋白质的高通量分析方面具有广泛的应用前景.基于量子点的稳定性和生物相容性,有可能通过标记不同颜色的量子点到不同的分子,观察它们在活细胞内的运动.     

Biofabrication of ZnS:Mn luminescent nanocrystals using histidine,hexahistidine, and His-tagged proteins: A comparison study

The ubiquitous hexahistidine purification tag has been used to conjugate proteins to the shell of CdSe:ZnS quantum dots (QDs) due to its affinity for surface-exposed Zn²⁺ ions but little attention has been paid to the potential of His-tagged proteins for mineralizing luminescent ZnS nanocrystals. Here, we compare the ability of free histidine, a His tag peptide, His-tagged thioredoxin (TrxA, a monomeric protein), and N- and C-terminally His-tagged versions of Hsp31 (a homodimeric protein) to support the synthesis of Mn-doped ZnS nanocrystals from aqueous precursors under mild conditions of pH (8.2) and temperature (37 °C). We find that: (1) it is possible to produce poor quality QDs when histidine is used at high (8 mM) concentration; (2) an increase in local histidine concentration through repetition of the amino acid as a His tag decreases the amount of needed reagent ≈10-fold and improves optical properties; (3) fusion of the same His tag to TrxA allows for ZnS:Mn QDs mineralization at micromolar concentrations; and (4) doubling the local hexahistidine concentration by exploiting Hsp31 dimerization further improves nanocrystal luminescence with the brightest particles obtained when His tags are spatially co-localized at the Hsp31 N-termini. Although hexahistidine tracts are not as efficient as combinatorially selected ZnS binding peptides at QD synthesis, it should be possible to use the large number of available His-tagged proteins and the synthesis approach described herein to produce luminescent nanoparticles whose protein shell carries a broad range of functions.     

Study on the interaction between 2‐mercaptoethanol,dimercaprol and CdSe quantum dots

The interactions between 2‐mercaptoethanol, dimercaprol and CdSe quantum dots (QDs) in organic media have been investigated by spectral methods. The results showed that the fluorescence (FL) emission of CdSe QDs gradually decreased, with a slight red‐shift, after adding thiols to CdSe QDs solutions. With the increase of the concentrations of thiols, the resonance light scattering (RLS) signal of CdSe QDs had been strongly enhanced in the wavelength range 300–500 nm, which was confirmed by the formation of larger CdSe QDs particles. The effect of thiols on the FL emission of CdSe QDs could be described by a Stern–Volmer‐type equation with the concentration ranges 1.0 × 10^–6–7.5 × 10^–4 mol/L for 2‐mercaptoethanol and 1.0 × 10^–7–2.5 × 10^–5 mol/L for dimercaprol. The possible mechanism of the interaction was proposed according to the results of UV‐vis absorption and micro‐Raman spectroscopy. The results indicated that FL quenching was mainly attributable to the exchange of the QDs surface molecules. Copyright © 2008 John Wiley & Sons, Ltd.     

Investigating Fluorescence Quenching of ZnS Quantum Dots by Silver Nanoparticles

Water dispersible zinc sulfide quantum dots (ZnS QDs) with an average diameter of 2.9 nm were synthesized in an environment friendly method using chitosan as stabilizing agent. These nanocrystals displayed characteristic absorption and emission spectra having an absorbance edge at 300 nm and emission maxima (λ _emission) at 427 nm. Citrate-capped silver nanoparticles (Ag NPs) of ca. 37-nm diameter were prepared by modified Turkevich process. The fluorescence of ZnS QDs was significantly quenched in presence of Ag NPs in a concentration-dependent manner with K _sv value of 9 × 10⁹ M⁻¹. The quenching mechanism was analyzed using Stern–Volmer plot which indicated mixed nature of quenching. Static mechanism was evident from the formation of electrostatic complex between positively charged ZnS QDs and negatively charged Ag NPs as confirmed by absorbance study. Due to excellent overlap between ZnS QDs emission and surface plasmon resonance band of Ag NPs, the role of energy transfer process as an additional quenching mechanism was investigated by time-resolved fluorescence measurements. Time-correlated single-photon counting study demonstrated decrease in average lifetime of ZnS QDs fluorescence in presence of Ag NPs. The corresponding F?rster distance for the present QD–NP pair was calculated to be 18.4 nm.     

High efficiency transport of quantum dots into plant roots with the aid of silwet L-77

Quantum dots (QDs) are a novel type of small, photostable and bright fluorophores that have been successfully applied to mammalian and human live cell imaging. In this study, highly dispersive water-soluble mercaptoacetic acid (MAA)-coated CdSe/ZnS QDs were synthesized, which were suitable for investigation as fluorescent probe labels. The treatment of maize seedling roots with QDs showed that the surfactant silwet L-77 aided the efficient transport of QDs into maize roots. Under a concentration ranging from 0.128 to 1.28 μM, QDs caused very low cytotoxicity on maize seed germination and root growth. The addition of mercuric chloride to the Hoagland solution resulted in a decrease of QD content in root tissues, and this decrease was reversed upon the addition of β-mercaptoethanol, which suggests that mercury-sensitive processes play a significant role in regulating QD flow in the maize root system. We speculate that the apoplastic pathway can contribute substantially to the total quantity of QDs reaching the stele. Therefore, based on this transport approach, MAA-coated QDs can be utilized for live imaging in plant systems to verify known physiological processes.     

Sulforaphane Protects the Liver against CdSe Quantum Dot-Induced Cytotoxicity

The potential cytotoxicity of cadmium selenide (CdSe) quantum dots (QDs) presents a barrier to their use in biomedical imaging or as diagnostic and therapeutic agents. Sulforaphane (SFN) is a chemoprotective compound derived from cruciferous vegetables which can up-regulate antioxidant enzymes and induce apoptosis and autophagy. This study reports the effects of SFN on CdSe QD-induced cytotoxicity in immortalised human hepatocytes and in the livers of mice. CdSe QDs induced dose-dependent cell death in hepatocytes with an IC₅₀ = 20.4 μM. Pre-treatment with SFN (5 μM) increased cell viability in response to CdSe QDs (20 μM) from 49.5 to 89.3%. SFN induced a pro-oxidant effect characterized by depletion of intracellular reduced glutathione during short term exposure (3–6 h), followed by up-regulation of antioxidant enzymes and glutathione levels at 24 h. SFN also caused Nrf2 translocation into the nucleus, up-regulation of antioxidant enzymes and autophagy. siRNA knockdown of Nrf2 suggests that the Nrf2 pathway plays a role in the protection against CdSe QD-induced cell death. Wortmannin inhibition of SFN-induced autophagy significantly suppressed the protective effect of SFN on CdSe QD-induced cell death. Moreover, the role of autophagy in SFN protection against CdSe QD-induced cell death was confirmed using mouse embryonic fibroblasts lacking ATG5. CdSe QDs caused significant liver damage in mice, and this was decreased by SFN treatment. In conclusion, SFN attenuated the cytotoxicity of CdSe QDs in both human hepatocytes and in the mouse liver, and this protection was associated with the induction of Nrf2 pathway and autophagy.     

Application of quantum dots as probes for correlative fluorescence, conventional, and energy-filtered transmission electron microscopy.

Luminescent semiconductor quantum dots (QDs) are a new class of fluorescent label with wide-ranging applications for cell imaging. The electron density and elemental composition of these materials permit the extension of their use as probes in conventional electron microscopy (TEM) and energy-filtered TEM (EFTEM). Here we illustrate the feasibility of using streptavidin-conjugated QDs as TEM tags by labeling a nuclear protein on cell sections and obtaining correlative fluorescence and TEM data. We also show that QD probes can be employed in conjunction with immunogold for co-localization of proteins at the ultrastructural level. Furthermore, by obtaining cadmium elemental maps of CdSe/ZnS QDs distributed on a nuclear structure, we demonstrate the potential of QDs for co-localization of multiple proteins when used in combination with EFTEM.     

Aqueous synthesis of highly stable CdTe/ZnS Core/Shell quantum dots for bioimaging

In this work, we report the synthesis, characterization and biological application of highly stable CdTe/ZnS (cadmium tellurite/zinc sulphide) Core/Shell (CS) quantum dots (QDs) capped with mercaptosuccinic acid (MSA). The CS QDs were synthesized using a simple one‐pot aqueous method. The synthesized CdTe/ZnS CS QDs were found to exhibit excellent stability even 100 days after preparation and also showed better photoluminescence quantum yield (PLQY) of about 50% compared with that of only CdTe QDs which was nearly 12%. The formation of the CdTe/ZnS CS was confirmed by high‐resolution transmission electron microscopy (HR‐TEM), and Fourier transform infra‐red (FTIR) and X‐ray diffraction (XRD) analyses. Further, on extending our study towards bioimaging of E. coli cells using the QDs samples, we found that CdTe/ZnS CS QDs showed better results compared with CdTe QDs.     

Spontaneous emission of semiconductor quantum dots in inverse opal SiO2 photonic crystals at different temperatures

The photoluminescence (PL) characteristics of CdSe quantum dots (QDs) infiltrated into inverse opal SiO₂ photonic crystals (PCs) are systemically studied. The special porous structure of inverse opal PCs enhanced the thermal exchange rate between the CdSe QDs and their surrounding environment. Finally, inverse opal SiO₂ PCs suppressed the nonlinear PL enhancement of CdSe QDs in PCs excited by a continuum laser and effectively modulated the PL characteristics of CdSe QDs in PCs at high temperatures in comparison with that of CdSe QDs out of PCs. The final results are of benefit in further understanding the role of inverse opal PCs on the PL characteristics of QDs. Copyright © 2015 John Wiley & Sons, Ltd.     

Nanoparticle targeting to neurons in a rat hippocampal slice culture model

We have previously shown that CdSe/ZnS core/shell luminescent semiconductor nanocrystals or QDs (quantum dots) coated with PEG [poly(ethylene glycol)]-appended DHLA (dihydrolipoic acid) can bind AcWG(Pal)VKIKKP₉GGH₆ (Palm1) through the histidine residues. The coating on the QD provides colloidal stability and this peptide complex uniquely allows the QDs to be taken up by cultured cells and readily exit the endosome into the soma. We now show that use of a polyampholyte coating [in which the neutral PEG is replaced by the negatively heterocharged CL4 (compact ligand)], results in the specific targeting of the palmitoylated peptide to neurons in mature rat hippocampal slice cultures. There was no noticeable uptake by astrocytes, oligodendrocytes or microglia (identified by immunocytochemistry), demonstrating neuronal specificity to the overall negatively charged CL4 coating. In addition, EM (electron microscopy) images confirm the endosomal egress ability of the Palm1 peptide by showing a much more disperse cytosolic distribution of the CL4 QDs conjugated to Palm1 compared with CL4 QDs alone. This suggests a novel and robust way of delivering neurotherapeutics to neurons.