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 DimensionsThe 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-stackFluorophore 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-stackCoordinates 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 resolutionAlexaFluor 488AlexaFluor 532Rhodamine 6GAlexaFluor 647Others.