Laura Martínez Maestro

Temperature sensing using fluorescent nanothermometers.

Acquiring the temperature of a single living cell is not a trivial task. In this paper, we devise a novel nanothermometer, capable of accurately determining the temperature of solutions as well as biological systems such as HeLa cancer cells. The nanothermometer is based on the temperature-sensitive fluorescence of NaYF(4):Er(3+),Yb(3+) nanoparticles, where the intensity ratio of the green fluorescence bands of the Er(3+) dopant ions ((2)H(11/2) --> (4)I(15/2) and (4)S(3/2) --> (4)I(15/2)) changes with temperature. The nanothermometers were first used to obtain thermal profiles created when heating a colloidal solution of NaYF(4):Er(3+),Yb(3+) nanoparticles in water using a pump-probe experiment. Following incubation of the nanoparticles with HeLa cervical cancer cells and their subsequent uptake, the fluorescent nanothermometers measured the internal temperature of the living cell from 25 degrees C to its thermally induced death at 45 degrees C.

CdSe Quantum Dots for Two-Photon Fluorescence Thermal Imaging

The technological development of quantum dots has ushered in a new era in fluorescence bioimaging, which was propelled with the advent of novel multiphoton fluorescence microscopes. Here, the potential use of CdSe quantum dots has been evaluated as fluorescent nanothermometers for two-photon fluorescence microscopy. In addition to the enhancement in spatial resolution inherent to any multiphoton excitation processes, two-photon (near-infrared) excitation leads to a temperature sensitivity of the emission intensity much higher than that achieved under one-photon (visible) excitation. The peak emission wavelength is also temperature sensitive, providing an additional approach for thermal imaging, which is particularly interesting for systems where nanoparticles are not homogeneously dispersed. On the basis of these superior thermal sensitivity properties of the two-photon excited fluorescence, we have demonstrated the ability of CdSe quantum dots to image a temperature gradient artificially created in a biocompatible fluid (phosphate-buffered saline) and also their ability to measure an intracellular temperature increase externally induced in a single living cell.

Subtissue Thermal Sensing Based on Neodymium-Doped LaF3 Nanoparticles

In this work, we report the multifunctional character of neodymium-doped LaF₃ core/shell nanoparticles. Because of the spectral overlap of the neodymium emission bands with the transparency windows of human tissues, these nanoparticles emerge as relevant subtissue optical probes. For neodymium contents optimizing the luminescence brightness of Nd³⁺:LaF₃ nanoparticles, subtissue penetration depths of several millimeters have been demonstrated. At the same time, it has been found that the infrared emission bands of Nd³⁺:LaF₃ nanoparticles show a remarkable thermal sensitivity, so that they can be advantageously used as luminescent nanothermometers for subtissue thermal sensing. This possibility has been demonstrated in this work: Nd³⁺:LaF₃ nanoparticles have been used to provide optical control over subtissue temperature in a single-beam plasmonic-mediated heating experiment. In this experiment, gold nanorods are used as nanoheaters while thermal reading is performed by the Nd³⁺:LaF₃ nanoparticles. The possibility of a real single-beam-controlled subtissue hyperthermia process is, therefore, pointed out.

CdTe Quantum Dots as Nanothermometers: Towards Highly Sensitive Thermal Imaging

2–4 ] Funda-mental studies concerning the nature and velocity of cellular dynamics also require an exact knowledge of the intracellular temperature, as it plays a relevant role in the rates at which cell division and enzyme reactions take place. Fundamental processes, such as adenosine triphosphate hydrolysis, can be also monitored by thermal imaging due to the presence of signifi cant thermogenesis.

Nanoparticles for highly efficient multiphoton fluorescence bioimaging

In this paper, we demonstrate for the first time that the new class of fluoride-based inorganic upconverting nanoparticles, NaYF4:Er3+, Yb3+, are the most efficient multiphoton excited fluorescent nanoparticles developed to date. The near-infrared-to-visible conversion efficiency of the aforementioned nanoparticles surpasses that of CdSe quantum dots and gold nanorods, which are the commercially available inorganic fluorescent nanoprobes presently used for multiphoton fluorescence bioimaging. The results presented here open new perspectives for the implementation of fluorescence tomography by multiphoton fluorescence imaging.

Fluorescent nanothermometers for intracellular thermal sensing

The importance of high-resolution intracellular thermal sensing and imaging in the field of modern biomedicine has boosted the development of novel nanosized fluorescent systems (fluorescent nanothermometers) as the next generation of probes for intracellular thermal sensing and imaging. This thermal mapping requires fluorescent nanothermometers with good biocompatibility and high thermal sensitivity in order to obtain submicrometric and subdegree spatial and thermal resolutions, respectively. This review describes the different nanosized systems used up to now for intracellular thermal sensing and imaging. We also include the later advances in molecular systems based on fluorescent proteins for thermal mapping. A critical overview of the state of the art and the future perspective is also included.

Fluorescent nanothermometers provide controlled plasmonic-mediated intracellular hyperthermia

AIM This article demonstrates how controlled hyperthermia at the cellular level can be achieved. MATERIALS & METHODS The method is based on the simultaneous intracellular incorporation of fluorescence nanothermometers (CdSe quantum dots) and metallic nanoheaters (gold nanorods). RESULTS Real-time spectral analysis of the quantum dot emission provides a detailed feedback about the intracellular thermal loading caused by gold nanorods excited at the plasmon frequency. Based on this approach, thermal dosimetry is assessed in such a way that the infrared laser (heating) power required to achieve catastrophic intracellular temperature increments in cancer cells is identified. CONCLUSIONS This pure optical method emerges as a new and promising guide for the development of infrared hyperthermia therapies with minimal invasiveness.

Quantum Dot Thermometry Evaluation of Geometry Dependent Heating Efficiency in Gold Nanoparticles

Quantum dot based thermometry, in combination with double beam confocal microscopy, was used to investigate the absorption/heating efficiency of gold nanoparticles with different morphologies (nanorods, nanocages, nanoshells, and nanostars), all of them with an intense localized surface plasmon resonance within the first biological window, at around 808 nm. The heating efficiency was found to be strongly dependent on the geometry of the nanostructure, with the largest values found for gold nanorods and long-edge gold nanostars, both of them with heating efficiencies close to 100%. Gold nanorods and nanocages were found to have the largest absorption cross section per unit mass among all the studied geometries, emerging as optimum photothermal agents with minimum metal loading for biosystems.

Quantum Dot‐Based Thermal Spectroscopy and Imaging of Optically Trapped Microspheres and Single Cells

Laser-induced thermal effects in optically trapped microspheres and single cells are investigated by quantum dot luminescence thermometry. Thermal spectroscopy has revealed a non-localized temperature distribution around the trap that extends over tens of micrometers, in agreement with previous theoretical models besides identifying water absorption as the most important heating source. The experimental results of thermal loading at a variety of wavelengths reveal that an optimum trapping wavelength exists for biological applications close to 820 nm. This is corroborated by a simultaneous analysis of the spectral dependence of cellular heating and damage in human lymphocytes during optical trapping. This quantum dot luminescence thermometry demonstrates that optical trapping with 820 nm laser radiation produces minimum intracellular heating, well below the cytotoxic level (43 °C), thus, avoiding cell damage.

Optical trapping of NaYF4:Er3+,Yb3+ upconverting fluorescent nanoparticles

We report on the first experimental observation of stable optical trapping of dielectric NaYF4:Er(3+),Yb(3+) upconverting fluorescent nanoparticles (~26 nm in diameter) using a continuous wave 980 nm single-beam laser. The laser serves both to optically trap and to excite visible luminescence from the nanoparticles. Sequential loading of individual nanoparticles into the trap is observed from the analysis of the emitted luminescence. We demonstrate that the trapping strength and the number of individual nanoparticles trapped are dictated by both the laser power and nanoparticle density. The possible contribution of thermal effects has been investigated by performing trapping experiment in both heavy water and into distilled water. For the case of heavy water, thermal gradients are negligible and optical forces dominate the trap loading behaviour. The results provide a promising path towards real three dimensional manipulation of single NaYF4:Er(3+),Yb(3+) nanoparticles for precise fluorescence sensing in biophotonics experiments.