Blanca del Rosal

1.3 μm emitting SrF2:Nd3+ nanoparticles for high contrast in vivo imaging in the second biological window

Novel approaches for high contrast, deep tissue, in vivo fluorescence biomedical imaging are based on infrared-emitting nanoparticles working in the so-called second biological window (1,000–1,400 nm). This allows for the acquisition of high resolution, deep tissue images due to the partial transparency of tissues in this particular spectral range. In addition, the optical excitation with low energy (infrared) photons also leads to a drastic reduction in the contribution of autofluorescence to the in vivo image. Nevertheless, as is demonstrated here, working solely in this biological window does not ensure a complete removal of autofluorescence as the specimen’s diet shows a remarkable infrared fluorescence that extends up to 1,100 nm. In this work, we show how the 1,340 nm emission band of Nd3+ ions embedded in SrF2 nanoparticles can be used to produce autofluorescence free, high contrast in vivo fluorescence images. It is also demonstrated that the complete removal of the food-related infrared autofluorescence is imperative for the development of reliable biodistribution studies.

Unveiling in Vivo Subcutaneous Thermal Dynamics by Infrared Luminescent Nanothermometers

The recent development of core/shell engineering of rare earth doped luminescent nanoparticles has ushered a new era in fluorescence thermal biosensing, allowing for the performance of minimally invasive experiments, not only in living cells but also in more challenging small animal models. Here, the potential use of active-core/active-shell Nd(3+)- and Yb(3+)-doped nanoparticles as subcutaneous thermal probes has been evaluated. These temperature nanoprobes operate in the infrared transparency window of biological tissues, enabling deep temperature sensing into animal bodies thanks to the temperature dependence of their emission spectra that leads to a ratiometric temperature readout. The ability of active-core/active-shell Nd(3+)- and Yb(3+)-doped nanoparticles for unveiling fundamental tissue properties in in vivo conditions was demonstrated by subcutaneous thermal relaxation monitoring through the injected core/shell nanoparticles. The reported results evidence the potential of infrared luminescence nanothermometry as a diagnosis tool at the small animal level.

Hybrid Nanostructures for High‐Sensitivity Luminescence Nanothermometry in the Second Biological Window

Hybrid nanostructures containing neodymium-doped nanoparticles and infrared-emitting quantum dots constitute highly sensitive luminescent thermometers operating in the second biological window. They demonstrate that accurate subtissue fluorescence thermal sensing is possible.

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.

Neodymium-doped nanoparticles for infrared fluorescence bioimaging: The role of the host

The spectroscopic properties of different infrared-emitting neodymium-doped nanoparticles (LaF3:Nd3+, SrF2:Nd3+, NaGdF4: Nd3+, NaYF4: Nd3+, KYF4: Nd3+, GdVO4: Nd3+, and Nd:YAG) have been systematically analyzed. A comparison of the spectral shapes of both emission and absorption spectra is presented, from which the relevant role played by the host matrix is evidenced. The lack of a “universal” optimum system for infrared bioimaging is discussed, as the specific bioimaging application and the experimental setup for infrared imaging determine the neodymium-doped nanoparticle to be preferentially used in each case.

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.

In vivo autofluorescence in the biological windows: the role of pigmentation.

Small animal deep-tissue fluorescence imaging in the second Biological Window (II-BW, 1000-1350 nm) is limited by the presence of undesirable infrared-excited, infrared-emitted (900-1700 nm) autofluorescence whose origin, spectral properties and dependence on strains is still unknown. In this work, the infrared autofluorescence and laser-induced whole body heating of five different mouse strains with distinct coat colors (black, grey, agouti, white and nude) has been systematically investigated. While neither the spectral properties nor the magnitude of organ autofluorescence vary significantly between mouse strains, the coat color has been found to strongly determine both the autofluorescence intensity as well as the laser-induced whole body heating. Results included in this work reveal mouse strain as a critical parameter that has to be seriously considered in the design and performance of small animal imaging experiments based on infrared-emitting fluorescent markers.

Overcoming Autofluorescence: Long-Lifetime Infrared Nanoparticles for Time-Gated In Vivo Imaging.

The always present and undesired contribution of autofluorescence is here completely avoided by combining a simple time gating technology with long lifetime neodymium doped infrared-emitting nanoparticles.

In Vivo Deep Tissue Fluorescence and Magnetic Imaging Employing Hybrid Nanostructures

Breakthroughs in nanotechnology have made it possible to integrate different nanoparticles in one single hybrid nanostructure (HNS), constituting multifunctional nanosized sensors, carriers, and probes with great potential in the life sciences. In addition, such nanostructures could also offer therapeutic capabilities to achieve a wider variety of multifunctionalities. In this work, the encapsulation of both magnetic and infrared emitting nanoparticles into a polymeric matrix leads to a magnetic-fluorescent HNS with multimodal magnetic-fluorescent imaging abilities. The magnetic-fluorescent HNS are capable of simultaneous magnetic resonance imaging and deep tissue infrared fluorescence imaging, overcoming the tissue penetration limits of classical visible-light based optical imaging as reported here in living mice. Additionally, their applicability for magnetic heating in potential hyperthermia treatments is assessed.

In Vivo Ischemia Detection by Luminescent Nanothermometers.

There is an urgent need to develop new diagnosis tools for real in vivo detection of first stages of ischemia for the early treatment of cardiovascular diseases and accidents. However, traditional approaches show low sensitivity and a limited penetration into tissues, so they are only applicable for the detection of surface lesions. Here, it is shown how the superior thermal sensing capabilities of near infrared-emitting quantum dots (NIR-QDs) can be efficiently used for in vivo detection of subcutaneous ischemic tissues. In particular, NIR-QDs make possible ischemia detection by high penetration transient thermometry studies in a murine ischemic hindlimb model. NIR-QDs nanothermometers are able to identify ischemic tissues by means of their faster thermal dynamics. In addition, they have shown to be capable of monitoring both the revascularization and damage recovery processes of ischemic tissues. This work demonstrates the applicability of fluorescence nanothermometry for ischemia detection and treatment, as well as a tool for early diagnosis of cardiovascular disease.