Abstract
The quality of fluorescence microscopy images is often impaired by the presence of sample induced optical aberrations. Adaptive optical elements such as deformable mirrors or spatial light modulators can be used to correct aberrations. However, previously reported techniques either require special sample preparation, or time consuming optimization procedures for the correction of static aberrations. This paper reports a technique for optical sectioning fluorescence microscopy capable of correcting dynamic aberrations in any fluorescent sample during the acquisition. This is achieved by implementing adaptive optics in a non conventional confocal microscopy setup, with multiple programmable confocal apertures, in which out of focus light can be separately detected, and used to optimize the correction performance with a sampling frequency an order of magnitude faster than the imaging rate of the system. The paper reports results comparing the correction performances to traditional image optimization algorithms, and demonstrates how the system can compensate for dynamic changes in the aberrations, such as those introduced during a focal stack acquisition though a thick sample.
Original language | English |
---|---|
Pages (from-to) | 949-959 |
Number of pages | 11 |
Journal | Optics Express |
Volume | 25 |
Issue number | 2 |
DOIs | |
Publication status | Published - 23 Jan 2017 |
Externally published | Yes |
Funding
European Research Council (ERC), (339681). Russian Ministry of Education ("5 in 100"). The research leading to these results has received funding from the European Research Council under the European Union's Seventh Framework Programme (FP7/2007-2013) / ERC grant agreement n. 339681 The work of G. Vdovin and O. Soloviev is partially funded by the program "5 in 100" of the Russian Ministry of Education, and by Flexible Optical B.V.. The authors would like to acknowledge the contributions of W.J.M. van Geest and C.J. Slinkman.