The SAFe nanoscopes can be incorporated onto any Olympus inverted microscope that features a camera port, making it an easy and flexible  nanoscopy solution. Our IX83 microscope is well suited for nanoscopy applications due to its stable frame, TruFocus™ Z-drift compensation module, and open structure.

SAFe nanoscopes can also be combined with our FV3000 confocal laser scanning microscope and the IXplore™ SpinSR super-resolution microscope system, enabling researchers to maximize their  imaging  capabilities with confocal microscopy, TIRFM, and SMLM in one system.

Raw image

Data processing

Image reconstruction

The ASTER illumination method offers a novel capability to exploit the entire FOV of sCMOS cameras for SMLM and TIRF imaging.

ASTER uses two galvanometer mirrors to control illumination at the sample plane. While the excitation beam keeps its position in the back focal plane (BFP), an angular rotation of a galvanometer induces a similar angle in the BFP, corresponding to a different position in the sample plane.

By applying specific patterns, such as raster scanning, ASTER can provide uniform excitation on tunable FOVs of up to 150 × 150 µm² for all excitation modes (EPI, HiLo, and TIRF).

Schematic of ASTER and the resulting illumation patterns.

Wide FOV TIRF Imaging with Homogenous Illumination

Olympus is a pioneer in the TIRF microscopy field, and our range of TIRF objectives is designed to provide tight control over the evanescent wave produced in TIRF imaging with magnifications ranging from 60X to 150X. The APON100XHOTIRF objective has the world’s highest NA of 1.7*, while the UPLAPO60XOHR and UPLAPO100XOHR are the world’s first plan apochromat objectives with a NA of 1.5.*

With Olympus’ optics and Abbelight’s ASTER illumination technology, users can achieve homogenous TIRF illumination over a wide FOV.

*_{As of November 2018. According to Olympus research.}

Cultured hippocampal neurons stained for spectrin cytoskeleton and imaged in TIRF microscopy mode. Homogeneous TIRF over the entire field of view of a Hamamatsu Fusion sCMOS camera (larger than the camera port size) was achieved using Abbelight ASTER technology. Sample courtesy of C; Leterrier, NeuroCytoLab, Marseille, and images by Adrien Mau, ISMO, Orsay.

U2OS cells stained for microtubules (alpha-Tubulin antibody) CF660, mitochondria (anti-TOMM20) CF680, and chromatin (EdU) AF647. Simultaneous multicolor 2D dSTORM with spectral demixing.

Multicolor Imaging Using Far-Red Spectral Demixing

Although 3D nanoscopy has revolutionized the fluorescence microscopy field by attaining unprecedented resolutions, multicolor imaging remains challenging in SMLM. This difficulty is due to several factors, including chromatic aberrations, the choice of optimal buffers, and the choice of single-molecule-compatible dyes.

To provide a solution, Abbelight has implemented spectral demixing for SMLM. By separating far-red dyes using a combination of the appropriate dichroic cube and ratiometric algorithms, this technique elegantly enables simultaneous multicolor imaging in SMLM.

In practical terms, it enables users to easily image in multicolor with one buffer, one laser, and one live acquisition.

Abbelight Product Brochure

Download the Abbelight Product brochure here (PDF, 5.3 MB)

SAFe Technical Brochure

Download the SAFe Technical Brochure here (PDF, 11.2 MB)

FV3000

Confocal Laser Scanning Microscope

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IXplore

Inverted Imaging Systems

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IXplore SpinSR

High-Content Screeinng

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