Visualization of axons and dendritic spines is crucial in neuroscience research.However,traditional microscopy is limited by diffraction-limited resolution and shallow imaging depth,making it difficult to study neuron...Visualization of axons and dendritic spines is crucial in neuroscience research.However,traditional microscopy is limited by diffraction-limited resolution and shallow imaging depth,making it difficult to study neuronal dynamics.Two-photon multifocal structured illumination microscopy(2P-MSIM)provides super-resolution imaging along with a reasonably good penetration,but it is vulnerable to optical aberrations in deep tissues.Herein we present a novel non-inertial scanning 2P-MSIM system incorporated with adaptive optics(AO)which allows for super-resolution imaging with effective aberration correction.Our strategy is designed to correct both laser and fluorescence paths simultaneously using a spatial light modulator and a deformable mirror respectively,providing better results than the individual path corrections.The successful implementation of adaptive optical two-photon multifocal structured illumination microscopy(AO 2P-MSIM)has allowed for the super-resolution imaging of neuronal structures in a mouse brain slice at great depths and dynamic morphological characteristics of zebrafish motoneurons in vivo.展开更多
基金National Key R&D Program of China(2021YFF0502900)National Natural Science Foundation of China(61975131,62175166,and 62127819)+1 种基金Shenzhen Key Laboratory of Photonics and Biophotonics(ZDSYS20210623092006020)Shenzhen Science and Technology Program(JCYJ20220818100202005,JCYJ20200109105411133).
文摘Visualization of axons and dendritic spines is crucial in neuroscience research.However,traditional microscopy is limited by diffraction-limited resolution and shallow imaging depth,making it difficult to study neuronal dynamics.Two-photon multifocal structured illumination microscopy(2P-MSIM)provides super-resolution imaging along with a reasonably good penetration,but it is vulnerable to optical aberrations in deep tissues.Herein we present a novel non-inertial scanning 2P-MSIM system incorporated with adaptive optics(AO)which allows for super-resolution imaging with effective aberration correction.Our strategy is designed to correct both laser and fluorescence paths simultaneously using a spatial light modulator and a deformable mirror respectively,providing better results than the individual path corrections.The successful implementation of adaptive optical two-photon multifocal structured illumination microscopy(AO 2P-MSIM)has allowed for the super-resolution imaging of neuronal structures in a mouse brain slice at great depths and dynamic morphological characteristics of zebrafish motoneurons in vivo.
