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

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Presentation transcript:

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; This requires switching of fluorochromes between on and off states followed by a sequential readout;

Super-resolution imaging The crucial step of dSTORM/GSD is to temporarily switch off the majority of fluorophores to allow the precise localization of single fluorophores.

Super-resolution as reconstructed z-stack Fluorophore in the focal plane: the image appears round; Below the focus: elongated in the vertical direction; Higher than the focus position: elongated in the horizontal direction.

Section localization scheme

Super-resolution as reconstructed z-stack Coordinates in z are assigned to a specific layer in a z-stack based on a calibration of the deformed PSF by the cylinder lens. Resolution around 70 nm can be achieved.

Evolution of resolution AlexaFluor 488 AlexaFluor 532 Rhodamine 6G AlexaFluor 647 Others.