Phasor-FLIM-guided unraveling of ATRA supramolecular organization in liposomal nanoformulations

Here we use fluorescence lifetime imaging microscopy (FLIM) to study the supramolecular organization of nanoencapsulated liposomal all- trans retinoic acid (ATRA), exploiting ATRA's intrinsic fluorescence as a source of signal and phasor transformation as a fit-free analytical approach to lifet...

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Bibliographic Details
Published in:Nanoscale 2023-12, Vol.15 (47), p.1985-199
Main Authors: Bernardi, Mario, Vernizzi, Michael, Baraldi, Laura, Balog, Sandor, Bassanetti, Irene, Sgarbi, Elisa, Fornasari, Luca, Arrigoni, Chiara, Cardarelli, Francesco
Format: Article
Language:English
Subjects:
Drugs
Fluorescence
Imaging techniques
Liposomes
Membranes
Microscopy, Fluorescence - methods
Phasors
Position measurement
Retinoic acid
Retinoids
Scattering, Small Angle
X-Ray Diffraction
X-ray scattering
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Summary:Here we use fluorescence lifetime imaging microscopy (FLIM) to study the supramolecular organization of nanoencapsulated liposomal all- trans retinoic acid (ATRA), exploiting ATRA's intrinsic fluorescence as a source of signal and phasor transformation as a fit-free analytical approach to lifetime data. Our non-invasive method is suitable for checking for the presence of a fraction of ATRA molecules interacting with liposomal membranes. The results are validated by independent small-angle X-ray scattering (SAXS) and nano-differential scanning calorimetry (NanoDSC) measurements, probing ATRA's putative position on the membrane and effect on membrane organization. Besides the insights on the specific case-study proposed, the present results confirm the effectiveness of Phasor-FLIM analysis in elucidating the nanoscale supramolecular organization of fluorescent drugs in pharmaceutical formulations. This underscores the importance of leveraging advanced imaging techniques to deepen our understanding and optimize drugs' performance in delivery applications. Fluorescence lifetime imaging microscopy (FLIM) is used to study the supramolecular organization of liposomal all- trans retinoic acid (ATRA), exploiting its intrinsic fluorescence as signal and phasor transformation as a fit-free analytical tool.
ISSN:2040-3364
2040-3372
DOI:10.1039/d3nr03420h