We evaluated cellular responses to polymer-treated gold nanorods, which were synthesized using the standard wet-chemistry method that utilizes hexadecyltrimethylammonium bromide (CTAB). The nanorod dispersions were coated with either polystyrene sulfonate (PSS) or polyethylene glycol (PEG). Two sizes of nanorods were tested, with optical responses peaking at 628 and 773 nm. The cells were from mammary adenocarcinoma (SKBR3), Chinese Hamster Ovary (CHO), mouse myoblast (C2C12) and Human Leukemia (HL60) cell lines. Their mitochondrial function following exposure to the nanorods were assessed using the MTS assay. We found PEGylated particles to have superior biocompatibility compared with PSS-coated nanorods, which showed substantial cytotoxicity. Electron microscopy showed no cellular uptake of PEGylated particles compared with their PSS counterparts. PEGylated gold nanorods also exhibited better dispersion stability in the presence of cell growth medium; PSS-coated rods tended to flocculate or cluster. In the case of the PSS particles, toxicity correlated with surface area across the two sizes of nanorods studied.

Gold nanorods (AuNR) can be tailored to possess an intense and narrow longitudinal plasmon (LP) absorption peak in the far-red to near-infrared wavelength region, where tissue is relatively transparent to light. This makes AuNRs excellent candidates as contrast agents for photoacoustic imaging, and as photothermal therapeutic agents. The favorable optical properties of AuNR which depend on the physical parameters of shape, size and plasmonic coupling effects, are required to be stable during use. We investigate the changes that are likely to occur in these physical parameters in the setting of photothermal therapeutics, and the influence that these changes have on the optical properties and the capacity to achieve target cell death. To this end we study 3 sets of interactions: pulsed light with AuNR, AuNR with cells, and pulsed light with cells incubated with AuNR. In the first situation we ascertain the threshold value of fluence required for photothermal melting or reshaping of AuNR to shorter AuNR or nanospheres, which results in drastic changes in optical properties. In the second situation when cells are exposed to antibody-conjugated AuNR, we observe using transmission electron microscopy (TEM) that the particles are closely packed and clustered inside vesicles in the cells. Using dark-field microscopy we show that plasmonic interactions between AuNRs in this situation causes blue-shifting of the LP absorption peak. As a consequence, no direct lethal damage to cells can be inflicted by laser irradiation at the LP peak. On the other hand, using irradiation at the transverse peak (TP) wavelength in the green, at comparative fluences, extensive cell death can be achieved. We attribute this behavior on the one hand to the photoreshaping of AuNR into spheres and on the other hand to clustering of AuNR inside cells. Both effects create sufficiently high optical absorption at 532 nm, which otherwise would have been present at the LP peak. We discuss implications of these finding on the application of these particles in biomedicine.

We have synthesized and characterized gold nanoparticles (spheres and rods) with optical extinction bands within the “optical imaging window.” The intense plasmon resonant driven absorption and scattering peaks of these nanoparticles make them suitable as contrast agents for optical imaging techniques. Further, we have conjugated these gold nanoparticles to a mouse monoclonal antibody specific to HER2 overexpressing SKBR3 breast carcinoma cells. The bioconjugation protocol uses noncovalent modes of binding based on a combination of electrostatic and hydrophobic interactions of the antibody and the gold surface. We discuss various aspects of the synthesis and bioconjugation protocols and the characterization results of the functionalized nanoparticles. Some proposed applications of these potential molecular probes in the field of biomedical imaging are also discussed.

The non-invasive quantification of total haemoglobin concentrations [tHb] is highly desired for the assessment of haematologic disorders in vulnerable patient groups, but invasive blood sampling is still the gold standard in current clinical practice. This work demonstrates the potential of visible-light spectroscopic optical coherence tomography (sOCT) for quantifying the [tHb] in human whole blood. To accurately quantify the [tHb] from the substantial optical attenuation by blood in the visible wavelength range, we used a combination of zero-delay acquisition and focus tracking that ensures optimal system sensitivity at any depth inside the sample. Subsequently, we developed an analysis model to adequately correct for the high scattering contribution by red blood cells to the sOCT signal. We validate our method and compare it to conventional sOCT (without focus tracking and zero-delay acquisition) through ex-vivo measurements on flowing human whole blood, with [tHb] values in the clinical range of 7–23 g/dL. For our method with optimized sensitivity, the measured and expected values correlate well (Pearson correlation coefficient = 0.89, p < 0.01), with a precision of 3.8 g/dL. This is a considerable improvement compared to conventional sOCT (Pearson correlation coefficient = 0.59, p = 0.16; precision of 9.1 g/dL).

Spatially confined measurements of bilirubin in tissue can be of great value for noninvasive bilirubin estimations during neonatal jaundice, as well as our understanding of the physiology behind bilirubin extravasation. This work shows the potential of spectroscopic visible-light optical coherence tomography (sOCT) for this purpose. At the bilirubin absorption peak around 460 nm, sOCT suffers from a strong signal decay with depth, which we overcome by optimizing our system sensitivity through a combination of zero-delay acquisition and focus tracking. In a phantom study, we demonstrate the quantification of bilirubin concentrations between 0 and 650 µM with only a 10% difference to the expected value, thereby covering the entire clinical pathophysiological range.

Rod shaped gold nanoparticles are synthesized using cetyltriammonium bromide (CTAB) as a major component of growth solutions. This surfactant is toxic to cells, but is at the moment unavoidable when monodisperse and high yield nanorods are to be synthesized. CTAB is found coating side walls of the nanoparticles and plays a role in maintaining colloidal stability. It may be displaced using thiolated PEG which is non-toxic to cells. Here we report on systematic studies of cell viability of such PEGylated nanorods on an SKBR3 cell-line using the MTS assay. These PEGylated particles are characterized using electron microscopy, optical spectroscopy and zeta potential measurements. It is expected that such treatment will be crucial in making nanorods compatible for in vivo biomedical applications.

With human milk being the most important source of infant nutrition, the protection and support of breastfeeding are essential from a global health perspective. Nevertheless, relatively few objective methods are available to investigate human milk composition and lactation physiology when a mother experiences breastfeeding problems. We argue that optics and photonics offer promising opportunities for this purpose. Any research activity within this new application field starts with a thorough understanding on how light interacts with human milk. Therefore, the aim of this study was to investigate the full set of optical properties for human milk and the biological variability therein. Using a novel approach that combines spatially resolved diffuse reflectance spectroscopy (SR-DRS) and spectroscopic optical coherence tomography (sOCT) between 450 and 650 nm, we quantified the absorption coefficient µ a , scattering coefficient µ s , reduced scattering coefficient µ s ', anisotropy g and backscattering coefficient µ b,NA of mature human milk from 14 participants released at different stages during a breastfeed (foremilk, bulk milk and hindmilk). Significant correlations were found between µ a , µ s , µ s ' and µ b,NA and the biochemically determined fat concentration per sample (R s = 0.38, R s = 0.77, R s = 0.80, R s = 0.44 respectively). We explained the observed variations in the optical properties of human milk using Mie theory and the biological variability in both the concentration and size distribution of milk fat globules. In conclusion, we have provided a full set of optical properties for human milk, which can hopefully serve as a starting point for future biophotonic studies on human milk and the milk containing lactating breast.

SummaryGold nanoparticles are rapidly emerging for use in biomedical applications. Characterization of the interaction and delivery of nanoparticles to cells through microscopy is important. Scanning electron microscopes have the intrinsic resolution to visualize gold nanoparticles on cells. A novel sample preparation protocol was developed to enable imaging of cells and gold nanoparticles with a conventional below lens scanning electron microscopes. The negative influence of 'charging' on the quality of scanning electron microscopes' images could be limited by deposition of biological cells on a conductive (gold) surface. The novel protocol enabled highresolution scanning electron microscopes' imaging of small clusters and individual gold nanoparticles on uncoated cell surfaces. Gold nanoparticles could be counted on cancer cells with automated routines.