Exploring Adaptive Optics for Controlled Aberration in STED and (STED-)FCS Microscopy

Session

Session 5 - Super-resolution and Nanoscale Imaging

Authors

Julius Trautmann (1), Christian Eggeling (1, 2)

Affiliations

1. Institute of Applied Optics and Biophysics, Friedrich Schiller University Jena
2. Leibniz Institute of Photonic Technology e.V. Jena

Keywords

Adaptive Optics 

AO

Fluorescence Correlation Spectroscopy

FCS

Stimulated Emission Depletion

STED

STED FCS

Deformable Mirror 

DM

Abstract text

We present the use of adaptive optics (AO) to induce controlled aberrations to impact two advanced microscopy techniques: Stimulated Emission Depletion (STED) imaging and Fluorescence Correlation Spectroscopy (FCS). FCS offers insights into molecular dynamics and interactions, while STED microscopy allows for super-resolved sub-diffraction imaging. Combining the strengths of both techniques, STED FCS emerges as a powerful approach that integrates STED imaging's high spatial resolution with FCS's temporal resolution, allowing for the investigation of dynamic processes with unprecedented precision at the nanoscale. However in biological experiments, delving deep into samples like tissue often introduces optical aberrations, compromising microscopy data quality within both imaging and spectroscopy. In recent years adaptive optics have become a remedy, employing active optical elements such as deformable mirrors (DMs) and spatial light modulators (SLMs)  

We here present our investigations on the influence of aberrations on STED microscopy imaging and (STED-)FCS measurements, where we have deliberately induced different aberrations and explored the outcome and possible corrections.