Correction of cell-induced optical aberrations in a fluorescence fluctuation microscope.
We describe the effect of optical aberrations on fluorescence fluctuations microscopy (FFM), when focusing through a single living cell. FFM measurements are performed in an aqueous fluorescent solution and prove to be a highly sensitive tool to assess the optical aberrations introduced by the cell. We demonstrate an adaptive optics (AO) system to remove the aberration-related bias in the FFM measurements. Our data show that AO is not only useful when imaging deep in tissues but also when performing FFM measurements through a single cellular layer. This work paves the way for the application of FFM to complex three-dimensional multicellular samples.
University of Grenoble 1/CNRS, LIPhy UMR 5588, Grenoble, France.
Charles-Edouard
CE
Leroux
Alexei
A
Grichine
Irène
I
Wang
Antoine
A
Delon
D000818:Animals; D016477:Artifacts; D002470:Cell Survival; D005347:Fibroblasts / Q000166:cytology*; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D055620:Optical Phenomena; D059010:Single-Cell Analysis / Q000379:methods*