Abstract
The emerging field of nanomedicine provides new approaches for the diagnosis and treatment of diseases, for symptom relief and for monitoring of disease progression. One route of realizing this approach is through carefully constructed nanoparticles. Due to the small size inherent to the nanoparticles a proper characterization is not trivial. This review highlights the application of time-resolved fluorescence spectroscopy and fluorescence lifetime imaging microscopy (FLIM) for the analysis of nanoparticles, covering aspects ranging from molecular properties to particle detection in tissue samples. The latter technique is particularly important as FLIM allows for distinguishing of target molecules from the autofluorescent background and, due to the environmental sensitivity of the fluorescence lifetime, also offers insights into the local environment of the nanoparticle or its interactions with other biomolecules. Thus, these techniques offer highly suitable tools in the fields of particle development, such as organic chemistry, and in the fields of particle application, such as in experimental dermatology or pharmaceutical research.
Highlights
We provide conformational dynamics of the polymer branches, as well as cargo partitioning and biomolecular insight on how the sensitivity of the fluorescence lifetime to the microenvironment can be used in interactions by using a classical organic fluorophore as a reporter group
Time-resolved fluorescence anisotropy measurements were used to study the model drug mimetic mimetic Nile Red loaded into core multishell (CMS) nanocarriers (CMS/NR) [61]
The results showed that the fluorescence decay of NR within the CMS nanocarrier contains fluorescence lifetime components found both for NR
Summary
Time-resolved fluorescence spectroscopy and imaging have been successfully used in elucidating structure and function of biological samples, e.g., proteins [4], and have the potential to provide an extremely powerful set of tools in the evaluation of dye-conjugated polymeric materials, nanoparticles, and nanocapsules [5,6]. In the study of nanoparticles time-resolved fluorescence measurements have not been used as much as steady-state approaches, such as the analysis of fluorescence emission spectra and steady-state anisotropy, even though they offer the possibility for a deeper understanding of the nanoparticle itself and the local environment as well as being able to provide information on loaded drug molecules. Recording the time-resolved fluorescence emission in a spatially resolved fashion on samples under a microscope, i.e., fluorescence lifetime imaging microscopy (FLIM), can provide localized information on fluorophores in cell and tissue samples. New developments in the analysis of fluorescence lifetime imaging microscopy data enable fast and reliable localization of target molecules, e.g., dye-nanoparticle conjugates, despite an autofluorescent background, allowing for the detection of fluorescently labeled nanoparticles in cellular systems and tissue samples [11,12,13,14,15,16]
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