Photoacoustic efect applied on model membranes and living cells: direct observation with multiphoton excitation microscopy and long-term viability analysis
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Galisteo González, FranciscoEditorial
Nature Research
Date
2020-01-15Referencia bibliográfica
Galisteo-González, F., Monasterio, B. G., Gil, D., Valle, M., & Goñi, F. M. (2020). Photoacoustic effect applied on model membranes and living cells: direct observation with multiphoton excitation microscopy and long-term viability analysis. Scientific reports, 10(1), 1-9. [https://doi.org/10.1038/s41598-019-56799-9]
Sponsorship
Spanish Ministry of Economy FEDER MINECO PGC2018-099857-B-I00; Basque Government IT1264-19 IT1270-19; University of Basque CountryAbstract
The photoacoustic efect is generated when a variable light interacts with a strongly light-absorbing
material. In water, it may produce hot bubbles and shock waves that could afect the integrity of
nearby cellular membranes, opening transient pores (photoporation). In this study, we have evaluated
the efect of pulsed laser-irradiated carbon nanoparticles (cNP) on model membranes and on Chinese
hamster ovary (CHO) cells. Fluorescence lifetime measurements of calcein-loaded liposomes support
the notion that the photoacoustic efect causes transient openings in membranes, allowing difusion
fuxes driven by gradient concentrations. With CHO cells, we have shown that this efect can induce
either intracellular delivery of calcein, or release of cellular compounds. The latter process has been
recorded live with multiphoton excitation microscopy during pulsed infrared laser irradiation. Calcein
loading and cell viability were assayed by fow cytometry, measuring necrotic cells as well as those in
early apoptosis. To further assess long-term cell recovery after the rather harsh treatment, cells were
reseeded and their behaviour recorded for 48h. These extended studies on cell viability show that
pulsed laser cNP photoporation may be considered an adequate intracellular delivery technique only if
employed with soft irradiation conditions (below 50mJ/cm2).