Open AccessFluorescent correlation spectroscopy measurements with adaptive optics in the intercellular space of spheroids
Author(s) -
Charles-Edouard Leroux,
Sylvain Monnier,
Irène Wang,
Giovanni Cappello,
Antoine Delon
Publication year - 2014
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.5.003730
Subject(s) - fluorescence correlation spectroscopy , optics , adaptive optics , brightness , diffusion , point spread function , biological system , metric (unit) , physics , fluorescence , materials science , biology , thermodynamics , operations management , economics
In this study we demonstrate the use of adaptive optics to correct the biasing effects of optical aberrations when measuring the dynamics of molecules diffusing between cells in multicellular spheroids. Our results indicate that, on average, adaptive optics leads to a reduction of the 3D size of the point spread function that is statistically significant in terms of measured number of molecules and diffusion time. The sensorless approach, which uses the molecular brightness as optimization metric, is validated in a complex, highly heterogeneous, biological environment. This work paves the way towards the design of accurate diffusion measurements of molecules in thick biological specimens.