A protocol for single‐source dual‐pulse stimulated emission depletion setup with Bessel modulation

Abstract

STimulated Emission Depletion (STED) microscopy attains super‐resolution in biological imaging beyond the diffraction limit. Here, we give a concise protocol to construct a dual‐pulse STED setup with one super‐continuum laser. Moreover, a flexible and dismountable Bessel modulation module is introduced for potential 2D‐stack STED imaging. Experiments and notices are introduced in detail, with discussion on some important check‐points for STED, such as detector saturation. Finally, the results validate the system working.

Raster-scanning Donut simplifies MINFLUX and provides alternative implement on other scanning-based microscopes

“…To reduce the engineered barrier and achieve fast construction, such as super-precision systems, RASTMIN is also an inspiration. With the popularization of the STED system review on spatial and temporal resolution 26 with the help of certain dyes 6 , 27 , moreover, the optically engineered STED protocol for self-building in its own lab 28 , the SSI-based localization system can quickly refit a prototype with performance close to MINFLUX for laboratories that have STED (especially stabilization is a commonly utilized path in observing GNs-samples’ sectioning PSF along both lateral and axial plane), or even more basically, confocal fluorescence microscope constructing experience. On this basis, do some avant-garde imaging tests or explorations, such as multi-photon multi-color MINFLUX 18 .…”