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Imaging tissue samples at sub-50 nm resolution in 3D by light microscopy
Light microscopy is traditionally limited by diffraction to about 250 nm resolution in the focal plane and more than 500 nm in depth. Super-resolution STED microscopy has overcome this diffraction limit but achieving sub-100 nm super-resolution in 3D in the middle of a tissue section has been impossible due to the optical aberrations the tissue introduces into the optical beam path. Introducing adaptive optics into an isoSTED microscope, an instrument that utilizes two opposing objectives for optimal 3D resolution, allowed the authors to correct for these aberrations. Using this instrument, they were able to obtain for the first time multi-color sub-50 nm 3D resolution images in samples as complex as Drosophila egg chambers and mouse brain tissue sections.
Source: Nature Methods