Luminescent single-walled carbon nanotube-sensitized europium nanoprobes for cellular imaging

Lanthanoid-based optical probes with excitation wavelengths in the ultra-violet (UV) range (300–325 nm) have been widely developed as imaging probes. Efficient cellular imaging requires that lanthanoid optical probes be excited at visible wavelengths, to avoid UV damage to cells. The efficacy of europium-catalyzed single-walled carbon nanotubes (Eu-SWCNTs), as visible nanoprobes for cellular imaging, is reported in this study. Confocal fluorescence microscopy images of breast cancer cells (SK-BR-3 and MCF-7) and normal cells (NIH 3T3), treated with Eu-SWCNT at 0.2 μg/mL concentration, showed bright red luminescence after excitation at 365 nm and 458 nm wavelengths. Cell viability analysis showed no cytotoxic effects after the incubation of cells with Eu-SWCNTs at this concentration. Eu-SWCNT uptake is via the endocytosis mechanism. Labeling efficiency, defined as the percentage of incubated cells that uptake Eu-SWCNT, was 95%–100% for all cell types. The average cellular uptake concentration was 6.68 ng Eu per cell. Intracellular localization was further corroborated by transmission electron microscopy and Raman microscopy. The results indicate that Eu-SWCNT shows potential as a novel cellular imaging probe, wherein SWCNT sensitizes Eu3+ ions to allow excitation at visible wavelengths, and stable time-resolved red emission. The ability to functionalize biomolecules on the exterior surface of Eu-SWCNT makes it an excellent candidate for targeted cellular imaging

Single-walled carbon nanotubes as a multimodal-thermoacoustic and photoacoustic-contrast agent

We have developed a novel carbon nanotube-based contrast agent for both thermoacoustic and photoacoustic tomography. In comparison to deionized water, single-walled carbon nanotubes exhibited more than twofold signal enhancement for thermoacoustic tomography at 3GHz. In comparison to blood, they exhibited more than sixfold signal enhancement for photoacoustic tomography at 1064nm wavelength. The large contrast enhancement of single-walled carbon nanotubes was further corroborated by tissue phantom imaging studies

Multiscale Photoacoustic Microscopy of Single-Walled Carbon Nanotube-Incorporated Tissue Engineering Scaffolds

Three-dimensional polymeric scaffolds provide structural support and function as substrates for cells and bioactive molecules necessary for tissue regeneration. Noninvasive real-time imaging of scaffolds and/or the process of tissue formation within the scaffold remains a challenge. Microcomputed tomography, the widely used technique to characterize polymeric scaffolds, shows poor contrast for scaffolds immersed in biological fluids, thereby limiting its utilities under physiological conditions. In this article, multiscale photoacoustic microscopy (PAM), consisting of both acoustic-resolution PAM (AR-PAM) and optical-resolution PAM (OR-PAM), was employed to image and characterize single-walled carbon-nanotube (SWNT)–incorporated poly(lactic-co-glycolic acid) polymer scaffolds immersed in biological buffer. SWNTs were incorporated to reinforce the mechanical properties of the scaffolds, and to enhance the photoacoustic signal from the scaffolds. By choosing excitation wavelengths of 570 and 638 nm, multiscale PAM could spectroscopically differentiate the photoacoustic signals generated from blood and from carbon-nanotube-incorporated scaffolds. OR-PAM, providing a fine lateral resolution of 2.6 μm with an adequate tissue penetration of 660 μm, successfully quantified the average porosity and pore size of the scaffolds to be 86.5%±1.2% and 153±15 μm in diameter, respectively. AR-PAM further extended the tissue penetration to 2 mm at the expense of lateral resolution (45 μm). Our results suggest that PAM is a promising tool for noninvasive real-time imaging and monitoring of tissue engineering scaffolds in vitro, and in vivo under physiological conditions

In vivo carbon nanotube-enhanced non-invasive photoacoustic mapping of the sentinel lymph node

Sentinel lymph node biopsy (SLNB), a less invasive alternative to axillary lymph node dissection (ALND), has become the standard of care for patients with clinically node-negative breast cancer. In SLNB, lymphatic mapping with radio-labeled sulfur colloid and/or blue dye helps identify the sentinel lymph node (SLN), which is most likely to contain metastatic breast cancer. Even though SLNB, using both methylene blue and radioactive tracers, has a high identification rate, it still relies on an invasive surgical procedure, with associated morbidity. In this study, we have demonstrated a non-invasive single-walled carbon nanotube (SWNT)-enhanced photoacoustic (PA) identification of SLN in a rat model. We have successfully imaged the SLN in vivo by PA imaging (793 nm laser source, 5 MHz ultrasonic detector) with high contrast-to-noise ratio (=89) and good resolution (~500 µm). The SWNTs also show a wideband optical absorption, generating PA signals over an excitation wavelength range of 740–820 nm. Thus, by varying the incident light wavelength to the near infrared region, where biological tissues (hemoglobin, tissue pigments, lipids and water) show low light absorption, the imaging depth is maximized. In the future, functionalization of the SWNTs with targeting groups should allow the molecular imaging of breast cancer

Novel breast cancer detection system combining both thermoacoustic (TA) and photoacoustic (PA) tomography using carbon nanotubes (CNTs) as a dual contrast agent

We report here a novel breast cancer scanner using microwave and light excitation and ultrasound detection. This combined thermoacoustic and photoacoustic tomography scanner is a nonionizing low cost system that can potentially provide high-resolution, dual contrast (microwave and light absorption) three dimensional images of the breast. Front breast compression will be used in this scanner to alleviate patient discomfort, experienced in side breast compression during traditional X-ray mammography. This scanner will use dry instead of gel ultrasonic coupling. We have also developed a carbon nanotube-based contrast agent for both thermoacoustic and photoacoustic imaging. In the future, targeted molecular photoacoustic and thermoacoustic imaging should be possible using this contrast agent

Single-walled carbon nanotubes as a multimodal-thermoacoustic and photoacoustic-contrast agent

We have developed a novel carbon nanotube-based contrast agent for both thermoacoustic and photoacoustic tomography. In comparison to deionized water, single-walled carbon nanotubes exhibited more than twofold signal enhancement for thermoacoustic tomography at 3GHz. In comparison to blood, they exhibited more than sixfold signal enhancement for photoacoustic tomography at 1064nm wavelength. The large contrast enhancement of single-walled carbon nanotubes was further corroborated by tissue phantom imaging studies

Dual-mode photoacoustic microscopy of carbon nanotube incorporated scaffolds in blood and biological tissues

Three-dimensional scaffolds provide physical support and an adjustable microenvironment to facilitate vascularization of ischemic tissues; however, in vivo imaging of scaffold functioning is still challenging. Micro-CT, the current frequentlyused imaging modality for scaffold characterization, provides poor contrast for wet scaffold, which limits its in vivo applications. In this paper, dual modes of photoacoustic microscopy (PAM), using acoustic resolution PAM (AR-PAM) and optical resolution PAM (OR-PAM), were employed for imaging scaffolds in blood as well as in chicken breast tissues. By choosing different wavelengths, 570 nm and 638 nm, we spectroscopically differentiated the photoacoustic signals generated from blood and from carbon nanotube incorporated scaffolds. The ex vivo experiments demonstrated a lateral resolution of 45 μm and a maximum penetration of ~2 mm for AR-PAM, and a lateral resolution of 3 μm and a maximum penetration of ~660 μm for OR-PAM. OR-PAM further quantified the average pore size of scaffolds to be 100-200 μm in diameter. Our results suggest that PAM is a promising tool for in vivo monitoring of scaffold-induced angiogenesis as well as the degradability of scaffolds themselves

Multiscale Photoacoustic Microscopy of Single-Walled Carbon Nanotube-Incorporated Tissue Engineering Scaffolds

Three-dimensional polymeric scaffolds provide structural support and function as substrates for cells and bioactive molecules necessary for tissue regeneration. Noninvasive real-time imaging of scaffolds and/or the process of tissue formation within the scaffold remains a challenge. Microcomputed tomography, the widely used technique to characterize polymeric scaffolds, shows poor contrast for scaffolds immersed in biological fluids, thereby limiting its utilities under physiological conditions. In this article, multiscale photoacoustic microscopy (PAM), consisting of both acoustic-resolution PAM (AR-PAM) and optical-resolution PAM (OR-PAM), was employed to image and characterize single-walled carbon-nanotube (SWNT)–incorporated poly(lactic-co-glycolic acid) polymer scaffolds immersed in biological buffer. SWNTs were incorporated to reinforce the mechanical properties of the scaffolds, and to enhance the photoacoustic signal from the scaffolds. By choosing excitation wavelengths of 570 and 638 nm, multiscale PAM could spectroscopically differentiate the photoacoustic signals generated from blood and from carbon-nanotube-incorporated scaffolds. OR-PAM, providing a fine lateral resolution of 2.6 μm with an adequate tissue penetration of 660 μm, successfully quantified the average porosity and pore size of the scaffolds to be 86.5%±1.2% and 153±15 μm in diameter, respectively. AR-PAM further extended the tissue penetration to 2 mm at the expense of lateral resolution (45 μm). Our results suggest that PAM is a promising tool for noninvasive real-time imaging and monitoring of tissue engineering scaffolds in vitro, and in vivo under physiological conditions