Aberration measurement and correction on a large field of view in fluorescence microscopy.

T Furieri,S Morara,G Calisesi,S Bonora,D Ancora,A Bassi

doi:10.1364/boe.441810

Abstract

The aberrations induced by the sample and/or by the sample holder limit the resolution of optical microscopes. Wavefront correction can be achieved using a deformable mirror with wavefront sensorless optimization algorithms but, despite the complexity of these systems, the level of correction is often limited to a small area in the field of view of the microscope. In this work, we present a plug and play module for aberration measurement and correction. The wavefront correction is performed through direct wavefront reconstruction using the spinning-pupil aberration measurement and controlling a deformable lens in closed loop. The lens corrects the aberrations in the center of the field of view, leaving residual aberrations at the margins, that are removed by anisoplanatic deconvolution. We present experimental results obtained in fluorescence microscopy, with a wide field and a light sheet fluorescence microscope. These results indicate that detection and correction over the full field of view can be achieved with a compact transmissive module placed in the detection path of the fluorescence microscope.

Highlights

Adaptive Optics is a technology used to improve image quality when the optical system is subject to phase aberrations
We present experimental results obtained in fluorescence microscopy, with a wide field and a light sheet fluorescence microscope
The adaptive lens (AL) is controlled by the spinningpupil aberration measurement (SPAM) with a closed loop, as in typical adaptive optics applications [19]

Summary

Introduction

Adaptive Optics is a technology used to improve image quality when the optical system is subject to phase aberrations. For example [10], [11], [12], propose methods that strongly simplify the system using an adaptive lens, instead of a deformable mirror, combined with optimization algorithms This has brought to promising results in microscopy and ophthalmic imaging [7], [13]. The detection is based on spinningpupil aberration measurement (SPAM) that reconstructs the wavefront phase in the different locations of the field of view [18] This device controls an adaptive lens based on multiple piezoelectric actuators [10], which is installed into the same chassis of the SPAM module, in a compact device. The remaining residual aberrations, still present in the other regions of the field of view, are removed by deconvolution This is possible because the corresponding point spread function is known, as the wavefront phase has been detected by SPAM over the entire sample

Methods

Results

Conclusion