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3D Localization Microscopy With Ground State Depletion ( GSD 3D)

3D Localization Microscopy With Ground State Depletion ( GSD 3D). Basic Principles of the GSD Super-Resolution Technology for Displaying Highly Resolved Cellular Structures in Three Dimensions. The exact position of a fluorochrome is based on the principle of temporal separation ;

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3D Localization Microscopy With Ground State Depletion ( GSD 3D)

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  1. 3D Localization Microscopy With Ground State Depletion (GSD 3D)

  2. Basic Principles of the GSD Super-Resolution Technology for Displaying Highly Resolved Cellular Structures in Three Dimensions • The exact position of a fluorochrome is based on the principle of temporal separation; • This requires switching of fluorochromesbetween on and off states followed by a sequential readout;

  3. Super-resolution imaging The crucial stepofdSTORM/GSD istotemporarily switch off themajorityoffluorophorestoallowthe precise localizationof single fluorophores.

  4. Super-resolution as reconstructed z-stack Fluorophorein the focal plane: the image appears round; Belowthefocus: elongated in the vertical direction; Higherthan the focus position: elongated in the horizontal direction.

  5. Sectionlocalizationscheme

  6. Super-resolution as reconstructed z-stack Coordinates in z are assignedto a specificlayer in a z-stackbasedon a calibrationofthedeformed PSF bythecylinderlens. Resolutionaround 70 nmcanbeachieved.

  7. Evolution of resolution AlexaFluor 488 AlexaFluor532 Rhodamine 6G AlexaFluor647 Others.

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