Introduction

The Abbelight SAFe 360 is an illumination system developed for 3D single molecule localization used in applications such as STORM, PALM, PAINT. Single Molecule Localization Microscopy (SMLM) utilizes sequential activation and temporal localization of photoswitchable fluorophores to create super resolution images. At any given moment, only an optically resolvable subset of fluorophores is excited and imaged, such that the position of each fluorophore can be determined with high precision. Subsequent iterations of this process with different subsets allows numerous fluorophores to be localized and a super-resolution image to be constructed.

The system is equipped with 405nm, 488nm, 532nm, 640nm lasers and is connected to an IX83 Olympus inverted microscope with and two pco.panda 4.2 sCMOS cameras. Using a dual camera system and an astigmatic lens, the astigmatic deformation of the PSF can be imaged separately in order to extract the relative axial position of a single particle relative to the focus plane of the objective.

The Abbelight SAFe 360 features a large homogeneous 150×150µm 2D field of view 15x15x15nm isotropic 3D localization precision, axial range of up to 5µm. Illumination modes: standard epifluorescence, Total Internal Reflection Fluorescence (TIRF) and Highly Inclined and Laminated Optical sheet (HiLo).

Acquisition & Analysis 

Acquisition is performed with the NEO_LiveImaging software which controls the instrument and acquisition parameters, live reconstruction of data, and features a variety of tools for analysis of clusters or single particle dynamics and export of images.

The NEO_analysis software is also installed for offline analysis on the CT Analysis computer which can be reserved in our BookItLab online reservation system.

New users

Please contact Maya Holdengreber, tel. 073-378-1106 to coordinate a meeting.

Reservations

For STORM or PALM experiments, the Abbelight SAFe 360 can be reserved through our BookItLab online reservation system (SAFe360 PRIMO).