Abbelight
Caterina Severi
Workshop Room 7
Biological imaging has successfully pierced the nanoscale in the recent years. Resolving the intricate interaction of multiple targets in a system, naturally represents the next step in the evolution of fluorescence nanoscopy. Among nanoscopy methods, Single molecule localization microscopy (SMLM) gives the best 3D spatial resolution and can offer inherently the largest FOV because of its origin from standard widefield imaging.
Several years ago, Abbelight co-developped with the group of Dr. Sandrine Lévêque-Fort, a new method of large uniform illumination called Adaptable Scanning for Tunable Excitation Regions (ASTER)1. Thanks to ASTER, which is integrated into the Abbelight SAFe imaging platform, it is now possible to acquire 150 x 150 µm² SMLM images, with nanoscopic resolution which is homogeneous across the whole FOV.
Two dimensional SMLM images are now straightforward to acquire. However, biological samples are three dimensional. Therefore, Abbelight SAFe imaging platform has developed and integrated the ultimate 3D method with different 3D super-localization strategies to get the best XYZ spatial resolution.
With this workshop, we would like to demonstrate the versatility of our SAFe imaging platform in acquiring large FOV 3D SMLM images. We will image several type of samples: COS7 cells, hippocampal rat neurons and others.
Schedule
15 min Introduction: SMLM, ASTER and 3D Strategies integrated into Abbelight setup (Presentation)
15 min Acquisition a large FOV 2D SMLM imaging
15 min Acquisition of Ultimate 3D SMLM images
15 min Summary of the workshop & Questions
1 A. Mau et al, Fast widefield scan provides tunable and uniform illumination optimizing super-resolution microscopy on large fields, Nat. Communication, 2021
Figure 1 a ASTER STORM imaging of COS-7 cells labeled for microtubules and an AF647-coupled secondary antibody, FOV size 200 µm × 200 µm, 20,000 frames at 20 fps. Extracted from [1]
Figure 2 : 3D Large FOV SMLM Images (Z color coded, 1 um imaging depth)