Time-resolved fluorescence determination of albumin using ZnGeO:Mn luminescence nanorods modified with polydopamine nanoparticles.

A novel time-resolved fluorescence (TRF) pobe is constructed to detect human serum albumin (HSA) by exploiting ZnGeO:Mn persistent luminescence nanorods (ZnGeO:Mn PLNRs) and polydopamine nanoparticles (PDA NPs). HSA-induced dynamic quenching leads to the fluorescence decrease of ZnGeO:Mn PLNRs, providing the basis for quantitative analysis of HSA. The excellent photo-thermal conversion performance of PDA NPs is helpful to the collision process between ZnGeO:Mn PLNRs and HSA, inducing significant improvement of sensitivity. HSA is quantified by measuring time-resolved fluorescence at 540 nm under excitation of 250-nm light. Under optimal conditions, HSA in the linear range 0.1-100 ng mL-1 are detected by this PDA-mediated ZnGeO:Mn probe with high sensitivity and selectivity, and the detection limit is 36 pg mL-1 (3σ/s). The RSD for the quantification of HSA (5 ng mL-1, n = 11) is 5.2%. The practicability of this TRF probe is confirmed by accurate monitoring HSA contents in urine samples, giving rise to satisfactory spiking recoveries of 96.2-106.0%.

Bioorthogonal SERS Nanotags as a Precision Theranostic Platform for in Vivo SERS Imaging and Cancer Photothermal Therapy.

Precise detection and effective treatment are crucial to prolong cancer patients' lives. Surface-enhanced Raman scattering (SERS) imaging coupled with photothermal therapy has been considered a precise and effective strategy for cancer theranostics. Nevertheless, Raman reporters employed in the literature usually possessed multiple shift peaks in the fingerprint region, which are overlapped with background signals from endogenous biological molecules. Herein, we fabricated a new kind of bioorthogonal Raman reporter and aptamer functionalized SERS nanotags. The SERS nanotags demonstrated a strong Raman signal at 2205 cm-1 in the biologically Raman-silent region and recognized MCF-7 breast cancer cells for Raman imaging with high specificity. Laser irradiation induced serious toxicity of MCF-7 cells due to the excellent photothermal capability of the SERS nanotags. After intravenous administration of the SERS nanotags, tumor Raman spectral detection and mapping in living mice were successfully achieved. Further in vivo antitumor experiments manifested that the aptamer-modified SERS nanotags significantly restrained tumor growth after laser irradiation with 99% inhibition rate and good biocompatibility. These results clearly revealed that the SERS nanotags could serve as a novel and precise theranostic platform for in vivo cancer diagnosis and photothermal therapy.

Multifunctional halloysite nanotubes for targeted delivery and controlled release of doxorubicin in-vitro and in-vivo studies.

The current state of cancer therapy encourages researchers to develop novel efficient nanocarriers. Halloysite nanotubes (HNTs) are good nanocarrier candidates due to their unique nanoscale (40-80 nm in diamter and 200-500 nm in length) and hollow lumen, as well as good biocompatibility and low cost. In our study, we prepared a type of folate-mediated targeting and redox-triggered anticancer drug delivery system, so that Doxorubicin (DOX) can be specifically transported to tumor sites due to the over-expressed folate-receptors on the surface of cancer cells. Furthermore, it can then be released by the reductive agent glutathione (GSH) in cancer cells where the content of GSH is nearly 103-fold higher than in the extracellular matrix. A series of methods have demonstrated that per-thiol-ß-cyclodextrin (ß-CD-(SH)7) was successfully combined with HNTs via a redox-responsive disulfide bond, and folic acid-polyethylene glycol-adamantane (FA-PEG-Ad) was immobilized on the HNTs through the strong complexation between ß-CD/Ad. In vitro studies indicated that the release rate of DOX raised sharply in dithiothreitol (DTT) reducing environment and the amount of released DOX reached 70% in 10 mM DTT within the first 10 h, while only 40% of DOX was released in phosphate buffer solution (PBS) even after 79 h. Furthermore, the targeted HNTs could be specifically endocytosed by over-expressed folate-receptor cancer cells and significantly accelerate the apoptosis of cancer cells compared to non-targeted HNTs. In vivo studies further verified that the targeted HNTs had the best therapeutic efficacy and no obvious side effects for tumor-bearing nude mice, while free DOX showed damaging effects on normal tissues. In summary, this novel nanocarrier system shows excellent potential for targeted delivery and controlled release of anticancer drugs and provides a potential platform for tumor therapy.

Two-Step Mechanism of Cellular Uptake of Cationic Gold Nanoparticles Modified by (16-Mercaptohexadecyl)trimethylammonium Bromide.

Cationic colloidal gold nanorods (GNRs) have a great potential as a theranostic tool for diverse medical applications. GNRs' properties such as cellular internalization and stability are determined by physicochemical characteristics of their surface coating. GNRs modified by (16-mercaptohexadecyl)trimethylammonium bromide (MTAB), MTABGNRs, show excellent cellular uptake. Despite their promise for biomedicine, however, relatively little is known about the cellular pathways that facilitate the uptake of GNRs, their subcellular fate and intracellular persistence. Here we studied the mechanism of cellular internalization and long-term fate of GNRs coated with MTAB, for which the synthesis was optimized to give higher yield, in various human cell types including normal diploid versus cancerous, and dividing versus nondividing (senescent) cells. The process of MTABGNRs internalization into their final destination in lysosomes proceeds in two steps: (1) fast passive adhesion to cell membrane mediated by sulfated proteoglycans occurring within minutes and (2) slower active transmembrane and intracellular transport of individual nanorods via clathrin-mediated endocytosis and of aggregated nanorods via macropinocytosis. The expression of sulfated proteoglycans was the major factor determining the extent of uptake by the respective cell types. Upon uptake into proliferating cells, MTABGNRs were diluted equally and relatively rapidly into daughter cells; however, in nondividing/senescent cells the loss of MTABGNRs was gradual and very modest, attributable mainly to exocytosis. Exocytosed MTABGNRs can again be internalized. These findings broaden our knowledge about cellular uptake of gold nanorods, a crucial prerequisite for future successful engineering of nanoparticles for biomedical applications such as photothermal cancer therapy or elimination of senescent cells as part of the emerging rejuvenation approach.

Rhodococcus aetherivorans BCP1 as cell factory for the production of intracellular tellurium nanorods under aerobic conditions.

BACKGROUND: Tellurite (TeO32-) is recognized as a toxic oxyanion to living organisms. However, mainly anaerobic or facultative-anaerobic microorganisms are able to tolerate and convert TeO32- into the less toxic and available form of elemental Tellurium (Te0), producing Te-deposits or Te-nanostructures. The use of TeO32--reducing bacteria can lead to the decontamination of polluted environments and the development of "green-synthesis" methods for the production of nanomaterials. In this study, the tolerance and the consumption of TeO32- have been investigated, along with the production and characterization of Te-nanorods by Rhodococcus aetherivorans BCP1 grown under aerobic conditions. RESULTS: Aerobically grown BCP1 cells showed high tolerance towards TeO32- with a minimal inhibitory concentration (MIC) of 2800 µg/mL (11.2 mM). TeO32- consumption has been evaluated exposing the BCP1 strain to either 100 or 500 µg/mL of K2TeO3 (unconditioned growth) or after re-inoculation in fresh medium with new addition of K2TeO3 (conditioned growth). A complete consumption of TeO32- at 100 µg/mL was observed under both growth conditions, although conditioned cells showed higher consumption rate. Unconditioned and conditioned BCP1 cells partially consumed TeO32- at 500 µg/mL. However, a greater TeO32- consumption was observed with conditioned cells. The production of intracellular, not aggregated and rod-shaped Te-nanostructures (TeNRs) was observed as a consequence of TeO32- reduction. Extracted TeNRs appear to be embedded in an organic surrounding material, as suggested by the chemical-physical characterization. Moreover, we observed longer TeNRs depending on either the concentration of precursor (100 or 500 µg/mL of K2TeO3) or the growth conditions (unconditioned or conditioned grown cells). CONCLUSIONS: Rhodococcus aetherivorans BCP1 is able to tolerate high concentrations of TeO32- during its growth under aerobic conditions. Moreover, compared to unconditioned BCP1 cells, TeO32- conditioned cells showed a higher oxyanion consumption rate (for 100 µg/mL of K2TeO3) or to consume greater amount of TeO32- (for 500 µg/mL of K2TeO3). TeO32- consumption by BCP1 cells led to the production of intracellular and not aggregated TeNRs embedded in an organic surrounding material. The high resistance of BCP1 to TeO32- along with its ability to produce Te-nanostructures supports the application of this microorganism as a possible eco-friendly nanofactory.

A temperature microsensor for measuring laser-induced heating in gold nanorods.

Measuring temperature is an extensively explored field of analysis, but measuring a temperature change in a nanoparticle is a new challenge. Here, a microsensor is configured to measure temperature changes in gold nanorods in solution upon laser irradiation. The device consists of a silicon wafer coated with silicon nitride in which a microfabricated resistance temperature detector was embedded and attached to a digital multimeter. A polydimethylsiloxane mold served as a microcontainer for the sample attached on top of the silicon membrane. This enables laser irradiation of the gold nanorods and subsequent measurement of temperature changes. The results showed a temperature increase of 8 to 10 °C and good correlation with theoretical calculations and bulk sample direct temperature measurements. These results demonstrate the suitability of this simple temperature microsensor for determining laser-induced heating profiles of metallic nanomaterials; such measurements will be essential for optimizing therapeutic and catalytic applications.

Gold nanoparticle silica nanopeapods.

A subnanometer gap-separated linear chain gold nanoparticle (AuNP) silica nanotube peapod (SNTP) was fabricated by self-assembly. The geometrical configurations of the AuNPs inside the SNTPs were managed in order to pose either a single-line or a double-line nanostructure by controlling the diameters of the AuNPs and the orifice in the silica nanotubes (SNTs). The AuNPs were internalized and self-assembled linearly inside the SNTs by capillary force using a repeated wet-dry process on a rocking plate. Transmission electron microscopy (TEM) images clearly indicated that numerous nanogap junctions with sub-1-nm distances were formed among AuNPs inside SNTs. Finite-dimension time domain (FDTD) calculations were performed to estimate the electric field enhancements. Polarization-dependent surface-enhanced Raman scattering (SERS) spectra of bifunctional aromatic linker p-mercaptobenzoic acid (p-MBA)-coated AuNP-embedded SNTs supported the linearly aligned nanogaps. We could demonstrate a silica wall-protected nanopeapod sensor with single nanotube sensitivity. SNTPs have potential application to intracellular pH sensors after endocytosis in mammalian cells for practical purposes. The TEM images indicated that the nanogaps were preserved inside the cellular constituents. SNTPs exhibited superior quality SERS spectra in vivo due to well-sustained nanogap junctions inside the SNTs, when compared to simply using AuNPs without any silica encapsulation. By using these SNTPs, a robust intracellular optical pH sensor could be developed with the advantage of the sustained nanogaps, due to silica wall-protection.

Library approach for reliable synthesis and properties of DNA-gold nanorod conjugates.

We developed a library-based approach to chemically stabilize cetyltrimethylammonium bromide (CTAB)-coated gold nanorods for the synthesis of polyvalent DNA-gold nanorod conjugates (DNA-AuNRs). Eleven chemical reagents were carefully chosen to constitute an additive library and screened by UV-vis spectroscopy to evaluate their stabilizing capability for the CTAB-coated AuNRs. Interestingly, 5-bromosalicylic acid (5-BrSA) was determined to most significantly stabilize the AuNRs by inducing additional adsorption of CTAB on the rod. Importantly, these stabilized AuNRs with 5-BrSA were conjugated with thiol DNA in an exceptionally reproducible and reliable method, which led to the systematic investigation of their cooperative assembly and disassembly properties under various conditions, including different types and lengths of the DNA sequences.

Phospholipid micelle-based magneto-plasmonic nanoformulation for magnetic field-directed, imaging-guided photo-induced cancer therapy.

We present a magnetoplasmonic nanoplatform combining gold nanorods (GNR) and iron-oxide nanoparticles within phospholipid-based polymeric nanomicelles (PGRFe). The gold nanorods exhibit plasmon resonance absorbance at near infrared wavelengths to enable photoacoustic imaging and photothermal therapy, while the Fe3O4 nanoparticles enable magnetophoretic control of the nanoformulation. The fabricated nanoformulation can be directed and concentrated by an external magnetic field, which provides enhancement of a photoacoustic signal. Application of an external field also leads to enhanced uptake of the magnetoplasmonic formulation by cancer cells in vitro. Under laser irradiation at the wavelength of the GNR absorption peak, the PGRFe formulation efficiently generates plasmonic nanobubbles within cancer cells, as visualized by confocal microscopy, causing cell destruction. The combined magnetic and plasmonic functionalities of the nanoplatform enable magnetic field-directed, imaging-guided, enhanced photo-induced cancer therapy. FROM THE CLINICAL EDITOR: In this study, a nano-formulation of gold nanorods and iron oxide nanoparticles is presented using a phospholipid micelle-based delivery system for magnetic field-directed and imaging-guided photo-induced cancer therapy. The gold nanorods enable photoacoustic imaging and photothermal therapy, while the Fe3O4 nanoparticles enable magnetophoretic control of the formulation. This and similar systems could enable more precise and efficient cancer therapy, hopefully in the near future, after additional testing.

Dual-modality single particle orientation and rotational tracking of intracellular transport of nanocargos.

The single particle orientation and rotational tracking (SPORT) technique was introduced recently to follow the rotational motion of plasmonic gold nanorod under a differential interference contrast (DIC) microscope. In biological studies, however, cellular activities usually involve a multiplicity of molecules; thus, tracking the motion of a single molecule/object is insufficient. Fluorescence-based techniques have long been used to follow the spatial and temporal distributions of biomolecules of interest thanks to the availability of multiplexing fluorescent probes. To know the type and number of molecules and the timing of their involvement in a biological process under investigation by SPORT, we constructed a dual-modality DIC/fluorescence microscope to simultaneously image fluorescently tagged biomolecules and plasmonic nanoprobes in living cells. With the dual-modality SPORT technique, the microtubule-based intracellular transport can be unambiguously identified while the dynamic orientation of nanometer-sized cargos can be monitored at video rate. Furthermore, the active transport on the microtubule can be easily separated from the diffusion before the nanocargo docks on the microtubule or after it undocks from the microtubule. The potential of dual-modality SPORT is demonstrated for shedding new light on unresolved questions in intracellular transport.

Sulfonated halloysite nanotubes as a novel cation exchange material for solid phase extraction of toxic pyrrolizidine alkaloids.

Pyrrolizidine alkaloids are phytochemicals, which present a highly toxic class of compounds in multiple food resources and are therefore a late-breaking topic in food safety. This study describes the first use of modified halloysite nanotubes as a novel solid material for solid phase extraction. As a result of a fast one-pot sulfonation of the cheap and non-toxic halloysite nanotubes, an efficient cation exchange phase has been prepared. After optimization of the solid phase extraction protocol, high extraction efficiencies and overall recoveries were obtained for a mixture of four pyrrolizidine alkaloid structures through UHPLC-MS/MS analysis with caffeine as the internal standard. Furthermore, the novel solid phase was used for the selective binding of the toxic pyrrolizidine alkaloids in a real-life honey sample, which itself is often contaminated with these compounds. In-house validation showed great extraction efficiencies up to 99.9% for senecionine with a lower limit for lycopsamine with 59.3%, which indicated high selectivity even in the presence of potential interfering compounds. Subsequently, overall recoveries up to 91.5% could be obtained for senecionine while the lowest value was reached for lycopsamine with 55.1%. Comparison with a commercial strong cation exchange tube procedure showed the high competitiveness of the novel solid phase with respect to overall performance. Only slight disadvantages regarding precision and repeatability with values under 5.7% and 11.6% could be observed. Therefore, sulfonated halloysite nanotubes present themselves as an easy to prepare, cheap and highly efficient novel cation exchange material for the selective solid phase extraction of toxic pyrrolizidine alkaloids in frequently contaminated real-life samples like honey.

MIL-101(Fe)@TiO2 nanotube composite material is used for the solid phase extraction of non-steroidal anti-inflammatory drugs under the synergy of multiple interactions.

Increasing the adsorption sites and effective interactions between sorbents and the targets can improve the solid-phase extraction (SPE) efficiency. Herein, based on the advantages of MOFs and TiO2 nanotubes (TiO2 NTs), an MIL-101(Fe)@TiO2 NT composite was prepared and applied to extract non-steroidal anti-inflammatory drugs (NSAIDs) from water samples coupled with high performance liquid chromatography (HPLC). Through characterization, it was established that MIL-101(Fe) was effectively composited on the surface and inside the TiO2 nanotubes, increasing effective adsorption sites. The obtained composite material well retains the structure and functional groups of the two original materials, and while retaining the original force with the target, it achieves a synergistic effect and produces more interactions with the target. Therefore, the extraction efficiency was greatly improved. The recovery efficiency reached 97.7-105.1% with an RSD of less than 6.71%, the detection limit was 0.1-0.2 µg L-1, and the linear range was 1-200 µg L-1 with a determination coefficient of 0.9972-0.9994. Owing to the stability of the two original materials, the composite material could be recycled and reused to extract NSAIDs up to 15 times without a loss of the recovery rate. Satisfactory results were obtained when it was used to extract NSAIDs from the Yellow River. These results indicate that the synthesized MIL-101(Fe)@TiO2 NT material is a promising sorbent to extract NSAIDs at trace concentrations with high efficiency and long lifetimes.

Zeolitic imidazolate framework-8 modified magnetic halloysite nanotube-based solid phase extraction for the analysis of carbamate pesticides by ultra-high performance liquid chromatography tandem mass spectrometry.

Zeolitic imidazolate framework-8 modified magnetic halloysite nanotube (MHNTs@ZIF-8) composites were synthesized and evaluated for the first time as an efficient sorbent for the magnetic solid-phase extraction (mSPE) of carbamate pesticides (CPs) from water samples. MHNTs were prepared by coprecipitation, and MHNTs@ZIF-8 composites were assembled in situ at room temperature. After characterization, MHNTs@ZIF-8 was used to extract pirimicarb, propoxur, carbaryl, isoprocarb and fenobucarb via π-π stacking interaction and hydrophobic interaction between the imidazole skeleton of ZIF-8 and benzene rings or benzene-like rings in CPs, as well as the hydrogen bond formed between O in CPs and H in ZIF-8. The effects of the amount of sorbent, ionic strength, type and volume of desorption solvent and adsorption/desorption time were investigated. Under optimum conditions, good linearity was obtained for the analysis of CPs by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) with R2 ≥ 0.9992. The limits of quantification range from 3 to 40 ng L-1 in water. Relative standard deviations (RSDs) were <7%, n = 5, within a batch and <9% among batches. The spiked recoveries were between 81 and 104%. The proposed method has been successfully applied to the determination of CPs in various water samples.

Synthesis and bioevaluation of ¹²5I-labeled gold nanorods.

A novel technique is described for monitoring the in vivo behavior of gold nanorods (GNRs) using γ-imaging. GNRs were radiolabeled using [¹²5I] sodium iodide in a simple and fast manner with high yield and without disturbing their optical properties. Radiolabeled GNRs were successfully visualized by radioisotope tagging, allowing longitudinal in vivo studies to be performed repeatedly in the same animal. The preliminary biodistribution study showed that PEGylated GNRs have much longer blood circulation times and clear out faster, while bare GNRs accumulate quickly in the liver after systematic administration. The highly efficient method reported here provides an extensively useful tool for guidance of the design and development of new gold nanoparticles as target-specific agents for both diagnostics and photothermal therapy.

Three-wavelength murine photoplethysmography for estimation of vascular gold nanorod concentration.

Nanoparticle-assisted photo-thermal (NAPT) ablation has become a new and attractive modality for the treatment of cancerous tumors. This therapy exploits the passive accumulation of intravenously delivered optically resonant metal nanoparticles into tumors, however, the circulating bioavailability of these particles is often unknown. We present a non-invasive optical device capable of monitoring the circulation of optically resonant gold nanorods. The device, referred to as a pulse photometer, uses the technique of multi-wavelength photoplethysmography. We simultaneously report the circulation of gold nanorods and oximetry for six hours post-injection in mice with no anesthesia and remove the probe when not collecting data. The instrument shows good agreement (R(2) = 0.903, n = 30) with ex vivo spectrophotometric analysis of blood samples. The real-time feedback provided has a strong potential for reducing variability and thus improving the efficacy of similar clinical therapies.

Nanorods versus nanospheres: a bifurcation mechanism revealed by principal component TEM analysis.

A quantitative analysis of object populations obtained by TEM images is performed for the classical scheme of aqueous seedless synthesis of nanorods. Using an effective way to represent nanoparticle size distributions, we unravel that spheres, usually considered to be a side-product, are in fact coming from a competing route during nanorod formation. The differentiation between spheres and rods appears above a critical size of 5 nm and is due to different growth rates between faces. The initial repartition of faces on nuclei or on the nanoparticles at the critical size can be the source for the final differentiation between globules and rods. The efficiency of the selection is strongly influenced by the production of the initial seeds and, in particular, by the amount of borohydride added in the present scheme.

A comparison study of detecting gold nanorods in living cells with confocal reflectance microscopy and two-photon fluorescence microscopy.

Two-photon fluorescence microscopy and confocal reflectance microscopy were compared to detect intracellular gold nanorods in rat basophilic leukaemia cells. The two-photon photoluminescence images of gold nanorods were acquired by an 800 nm fs laser with the power of milliwatts. The advantages of the obtained two-photon photoluminescence images are high spatial resolution and reduced background. However, a remarkable photothermal effect on cells was seen after 30 times continuous scanning of the femto-second laser, potentially affecting the subcellular localization pattern of the nanorods. In the case of confocal reflectance microscopy the images of gold nanorods can be obtained with the power of light source as low as microwatts, thus avoiding the photothermal effect, but the resolution of such images is reduced. We have noted that confocal reflectance images of cellular gold nanorods achieved with 50 microW 800 nm fs have a relatively poor resolution, whereas the 50 microW 488 nm CW laser can acquire reasonably satisfactory 3D reflectance images with improved resolution because of its shorter wavelength. Therefore, confocal reflectance microscopy may also be a suitable means to image intracellular gold nanorods with the advantage of reduced photothermal effect.

Characterization of gold nanorods in vivo by integrated analytical techniques: their uptake, retention, and chemical forms.

Integrated analytical techniques were used to study the tissue distribution and structural information of gold nanorods (Au NRs) in Sprague-Dawley rats through tail intravenous injection. Before in vivo experiments were conducted, careful characterization of Au NRs was performed. The zeta potential proved that adsorption of bovine serum albumin on Au NRs turned the surface charges from positive to negative as in an in vitro simulation. The biodistribution of Au NRs was investigated quantitatively by inductively coupled plasma mass spectrometry at different time points after injection. As target tissues, both liver and spleen were chosen to further demonstrate the intracellular localization of Au NRs by the combination of transmission electron microscopy and energy-dispersive X-ray spectroscopy. Moreover, synchrotron-radiation-based X-ray absorption spectroscopy was employed and it was observed that long-term retention of Au NRs in liver and spleen did not induce obvious changes in the oxidation states of gold. Therefore, the present systematic method can provide important information about the fates of Au NRs in vivo and can also be extended to study the biological effects of other metallic nanomaterials in the future.

Polyvalent Nanoobjects for Precision Diagnostics.

As our ability to synthesize and modify nanoobjects has improved, efforts to explore nanotechnology for diagnostic purposes have gained momentum. The variety of nanoobjects, especially those with polyvalent properties, displays a wide range of practical and unique properties well suited for applications in various diagnostics. This review briefly covers the broad scope of multivalent nanoobjects and their use in diagnostics, ranging from ex vivo assays and biosensors to in vivo imaging. The nanoobjects discussed here include silica nanoparticles, gold nanoparticles, quantum dots, carbon dots, fullerenes, polymers, dendrimers, liposomes, nanowires, and nanotubes. In this review, we describe recent reports of novel applications of these various nanoobjects, particularly as polyvalent entities designed for diagnostics.

Molecular fingerprinting of nanoparticles in complex media with non-contact photoacoustics: beyond the light scattering limit.

Optical instruments can probe physical systems even to the level of individual molecules. In particular, every molecule, solution, and structure such as a living cell has a unique absorption spectrum representing a molecular fingerprint. This spectrum can help identify a particular molecule from others or quantify its concentration; however, scattering limits molecular fingerprinting within a complex compound and must be overcome. Here, we present a new, non-contact photoacoustic (PA)-based method that can almost completely remove the influence of background light scattering on absorption measurements in heterogeneous highly scattering solutions and, furthermore, separate the intrinsic absorption of nanoscale objects from their scattering. In particular, we measure pure absorption spectra for solutions of gold nanorods (GNRs) as an example of a plasmonic agent and show that these spectra differ from the extinction measured with conventional UV-VIS spectrophotometry. Finally, we show how the original GNR absorption changes when nanoparticles are internalized by cells.