Aberration correction in stimulated emission depletion microscopy to increase imaging depth in living brain tissue

Significance: Stimulated emission depletion (STED) microscopy enables nanoscale imaging of live samples, but it requires a specific spatial beam shaping that is highly sensitive to optical aberrations, limiting its depth penetration. Therefore, there need for methods reduce aberrations and improve the resolution STED inside thick biological tissue. Aim: The aim our work was develop validate method based on adaptive optics achieve an priori correction spherical as function depth. Approach: We first measured in phantom sample gold fluorescent nanoparticles suspended agarose gel with refractive index closely matching living brain then used light modulator apply corrective phase shifts this calibration approach by neurons slices. Results: After quantifying we demonstrated corrections can substantially increase image quality Specifically, could measure structures small 80 nm at 90 μm tissue obtain 60% signal after correction. Conclusion: propose simple robust calibrate compensate distortions profile introduced increasing offers significant improvements performance cellular