Combined two-photon luminescence microscopy and OCT for macrophage detection in the hypercholesterolemic rabbit aorta using plasmonic gold nanorose.

BACKGROUND AND OBJECTIVES: The macrophage is an important early cellular marker related to risk of future rupture of atherosclerotic plaques. Two-channel two-photon luminescence (TPL) microscopy combined with optical coherence tomography (OCT) was used to detect, and further characterize the distribution of aorta-based macrophages using plasmonic gold nanorose as an imaging contrast agent. STUDY DESIGN/MATERIALS AND METHODS: Nanorose uptake by macrophages was identified by TPL microscopy in macrophage cell culture. Ex vivo aorta segments (8 × 8 × 2 mm(3) ) rich in macrophages from a rabbit model of aorta inflammation were imaged by TPL microscopy in combination with OCT. Aorta histological sections (5 µm in thickness) were also imaged by TPL microscopy. RESULTS: Merged two-channel TPL images showed the lateral and depth distribution of nanorose-loaded macrophages (confirmed by RAM-11 stain) and other aorta components (e.g., elastin fiber and lipid droplet), suggesting that nanorose-loaded macrophages are diffusively distributed and mostly detected superficially within 20 µm from the luminal surface of the aorta. Moreover, OCT images depicted detailed surface structure of the diseased aorta. CONCLUSIONS: Results suggest that TPL microscopy combined with OCT can simultaneously reveal macrophage distribution with respect to aorta surface structure, which has the potential to detect vulnerable plaques and monitor plaque-based macrophages overtime during cardiovascular interventions.

Effect on blood glucose monitoring of skin pressure exerted by an optical coherence tomography probe.

We proposed to use optical coherence tomography (OCT) for continuous noninvasive blood glucose monitoring, and recently we significantly improved the sensitivity of this technique. The accuracy of OCT glucose monitoring is limited by several factors, including variation of tissue pressure exerted by the OCT probe. We studied the influence of high (>10 kPa) and low (<1 kPa) pressure levels on OCT blood glucose monitoring. We showed that controlling external pressure to <1 kPa substantially improved the accuracy and reproducibility of OCT-based glucose monitoring.

Influence of osmolytes on in vivo glucose monitoring using optical coherence tomography.

Diabetes mellitus and its complications are the third leading cause of death in the world, exceeded only by cardiovascular disease and cancer. Tighter monitoring and control of blood glucose could minimize complications associated with diabetes. Recently, optical coherence tomography (OCT) for noninvasive glucose monitoring was proposed and tested in vivo. The aim of this work was to investigate the influence of changes in blood glucose concentration ([glu]) and sodium concentration ([Na+]) on the OCT signal. We also investigated the influence of other important analytes on the sensitivity of glucose monitoring with OCT. The experiments were carried out in anesthetized female pigs. The OCT images were acquired continuously from skin, while [glu] and [Na+] were experimentally varied within their physiological ranges. Correlations of the OCT signal slope with [glu] and [Na+] were studied at different tissue depths. The tissue area probed with OCT was marked and cut for histological examination. The correlation of blood [glu] and [Na+] with the OCT signal slope was observed in separate tissue layers. On average, equimolar changes in [glu] produced 2.26 +/- 1.15 greater alterations of the OCT signal slope than changes in [Na+]. Variation of concentrations of other analytes did not influence the OCT signal slope. The influence of [Na+] on relative changes in the OCT signal slope was generally less than [glu]-induced changes. OCT is a promising method for noninvasive glucose monitoring because of its ability to track the influence of changing [glu] on individual tissue layers.

In vivo study of glucose-induced changes in skin properties assessed with optical coherence tomography.

Recently, our in vivo studies demonstrated a strong correlation between blood glucose concentration and the slope of the optical coherence tomography (OCT) signal when the probing beam was scanned over a straight line. To improve the sensitivity of OCT for blood glucose monitoring, two-dimensional (2D) lateral scanning of the OCT probing beam was proposed. Depth-dependent changes in pig skin properties with variation of blood glucose concentration were revealed due to significant suppression of speckle noise and motion artefacts in 2D scanning mode. The correlation coefficient of the OCT signal slope with blood glucose concentration varied periodically in the range from -0.9 to +0.9 depending on depth. The period of variation of the correlation coefficient was 100-150 microm that corresponded to the distance between neighbour collagen bundles. We also observed a decrease of skin thickness by 10 +/- 7.5 microm with an increase of blood glucose concentration by 277 +/- 56 mg dl(-1). Mechanisms of glucose-induced changes in skin properties owing to tissue layer shift caused by dehydration associated with the glucose osmotic effect were considered.

Use of near-infrared luminescent gold nanoclusters for detection of macrophages.

We determined the effect of aggregation and coating thickness of gold on the luminescence of nanoparticles engulfed by macrophages and in gelatin phantoms. Thin gold-coated iron oxide nanoclusters (nanoroses) have been developed to target macrophages to provide contrast enhancement for near-infrared optical imaging applications. We compare the brightness of nanoroses luminescent emissions in response to 635 nm laser excitation to other nanoparticles including nanoshells, nanorods, and Cy5 conjugated iron oxide nanoparticles. Luminescent properties of all these nanoparticles were investigated in monomeric and aggregated form in gelatin phantoms and primary macrophage cell cultures using confocal microscopy. Aggregation of the gold nanoparticles increased luminescence emission and correlated with increased surface mass of gold per nanoparticle (nanoshells 37 ± 14.30 × 10(-3) brightness with 1.23 × 10(-4) wt of gold (g)/nanoparticle versus original nanorose 1.45 ± 0.37 × 10(-3) with 2.10 × 10(-16) wt of gold/nanoparticle, p<0.05). Nanoshells showed greater luminescent intensity than original nanoroses or Cy5 conjugated iron oxide nanoparticles when compared as nanoparticles per macrophage (38 ± 10 versus 11 ± 2.8 versus 17 ± 6.5, p<0.05, respectively, ANOVA), but showed relatively poor macrophage uptake (1025 ± 128 versus 7549 ± 236 versus 96,000 nanoparticles/cell, p<0.05, student t-test nanoshells versus nanoroses). Enhancement of gold fluorescent emissions by nanoparticles can be achieved by reducing the thickness of the gold coating, by clustering the gold on the surface of the nanoparticles (nanoshells), and by clustering the gold nanoparticles themselves.

Dual-wavelength multifrequency photothermal wave imaging combined with optical coherence tomography for macrophage and lipid detection in atherosclerotic plaques using gold nanoparticles.

The objective of this study was to assess the ability of combined photothermal wave (PTW) imaging and optical coherence tomography (OCT) to detect, and further characterize the distribution of macrophages (having taken up plasmonic gold nanorose as a contrast agent) and lipid deposits in atherosclerotic plaques. Aortas with atherosclerotic plaques were harvested from nine male New Zealand white rabbits divided into nanorose- and saline-injected groups and were imaged by dual-wavelength (800 and 1210 nm) multifrequency (0.1, 1 and 4 Hz) PTW imaging in combination with OCT. Amplitude PTW images suggest that lateral and depth distribution of nanorose-loaded macrophages (confirmed by two-photon luminescence microscopy and RAM-11 macrophage stain) and lipid deposits can be identified at selected modulation frequencies. Radiometric temperature increase and modulation amplitude of superficial nanoroses in response to 4 Hz laser irradiation (800 nm) were significantly higher than native plaque (P<0.001). Amplitude PTW images (4 Hz) were merged into a coregistered OCT image, suggesting that superficial nanorose-loaded macrophages are distributed at shoulders on the upstream side of atherosclerotic plaques (P<0.001) at edges of lipid deposits. Results suggest that combined PTW-OCT imaging can simultaneously reveal plaque structure and composition, permitting characterization of nanorose-loaded macrophages and lipid deposits in atherosclerotic plaques.

Correlation between optical coherence tomography images and histology of pigskin.

Noninvasive imaging techniques such as optical coherence tomography (OCT) are being widely used for early diagnostics of a variety of pathologies. Traditional tissue preparation for histological evaluation alters the dimensions of histological images such that differences between the dimensions of histologic and OCT images can be as great as 2.5-fold. This discrepancy complicates quantitative interpretation of OCT images. Here we used a steel ring sewn to the pigskin to avoid tissue deformation due to excision and fixation and used a surgical suture as a fixed tissue marker. With these techniques, we achieved good correlation between OCT and histological images with differences of less than 10%. The resulting method significantly enhances the diagnostic capabilities of the OCT technique.

Prediction capability of optical coherence tomography for blood glucose concentration monitoring.

BACKGROUND: Optical coherence tomography (OCT) has been shown to be a promising optical approach to noninvasive monitoring of blood glucose concentration because of its capability of probing optical properties at different depths in tissue with high resolution. This article investigates the capability of OCT to predict changes in blood glucose concentration. METHODS: We varied blood glucose concentration in the physiological range in three sets of experiments. In the first set, we investigated large variations of blood glucose concentration ( approximately 400 mg/dl) and used 2100 OCT A scans for signal averaging. In the second set, we varied blood glucose concentration by approximately 200 mg/dl and used 8400 A scans for signal averaging. In the third set, we improved OCT blood glucose monitoring by increasing and controlling skin temperature under the OCT probe. In this set of experiments we increased the glucose concentration by approximately 300 mg/dl and used 4200 A scans for averaging. RESULTS: The predicted glucose concentrations in the first two sets were lower than actual glucose concentration by 10-20% (the mean shift), while the heating and temperature control in the third set of experiments reduced the mean shift down to 1.5%. Therefore, the mean shift was reduced substantially by tissue heating and temperature control. However, it did not depend on the number of A scans to be averaged. In contrast, the uncertainty in OCT prediction of glucose concentration (the standard deviation) did not depend on heating and temperature control, but was reduced substantially from 56 to 24 mg/dl by increasing the number of A scans from 2100 to 8400, respectively. CONCLUSION: These results suggest that the accuracy of OCT-based glucose monitoring is approaching that of standard invasive and minimally invasive techniques.

In vivo visualization of Tradescantia leaf tissue and monitoring the physiological and morphological states under different water supply conditions using optical coherence tomography.

The optical coherence tomography (OCT) capabilities of plants were evaluated using leaves of Tradescantia pallida (Rose) D. Hunt. The internal structure of the leaf tissues was visualized in vivo and the physiological and morphological states of the tissues under different water supply conditions were monitored using OCT. The OCT technique provides non-invasive two-dimensional images directly on intact plants. The acquisition time of a two-dimensional image with a size of 200x200 pixels and a spatial resolution of 15 microm is 1-3 s. It was shown that OCT is a useful tool for monitoring the physiological and morphological states of plant tissues supplied with varying amounts of water and under the influence of different chemical factors.