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.

Heating efficiency of multi-walled carbon nanotubes in the first and second biological windows.

Quantum dot based-thermometry, in combination with double beam confocal microscopy and infrared thermal imaging, has been used to investigate the heating efficiency of multi-walled carbon nanotubes (MWCNTs) under optical excitation within the first (808 nm) and second (1090 nm) biological windows as well as in the spectral region separating them (980 nm). It has been found that for the three excitation wavelengths the heating efficiency of MWCNTs (10 nm in diameter and 1.5 µm in length) is close to 50%. Despite this "flat" heating efficiency, we have found that the excitation wavelength is, indeed, critical during in vivo experiments due to the spectral dependence of both tissue absorption and scattering coefficients. It has been concluded that efficiency and selectivity of in vivo photothermal treatments based on MWCNTs are simultaneously optimized when laser irradiation lies within the first or second biological window.

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.

NIR-to-NIR two-photon excited CaF2:Tm3+,Yb3+ nanoparticles: multifunctional nanoprobes for highly penetrating fluorescence bio-imaging.

In this study, we report on the remarkable two-photon excited fluorescence efficiency in the "biological window" of CaF(2):Tm(3+),Yb(3+) nanoparticles. On the basis of the strong Tm(3+) ion emission (at around 800 nm), tissue penetration depths as large as 2 mm have been demonstrated, which are more than 4 times those achievable based on the visible emissions in comparable CaF(2):Er(3+),Yb(3+) nanoparticles. The outstanding penetration depth, together with the fluorescence thermal sensitivity demonstrated here, makes CaF(2):Tm(3+),Yb(3+) nanoparticles ideal candidates as multifunctional nanoprobes for high contrast and highly penetrating in vivo fluorescence imaging applications.

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.

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.