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ASTER Technology for High Localization Precision over a Wide Field of View

The SAFe nanoscopes all share the same unique excitation system, which is based on the Adaptable Scanning for Tunable Excitation Regions (ASTER) technology.1 ASTER generates homogenous illumination in TIRF, HiLo, and EPI modes while performing SMLM modalities such as PALM, STORM, or PAINT with a localization precision reaching 10–15 nm in 3D on a field of view (FOV) of 150 x 150 µm2.

Schematic of ASTER and the resulting illumation patterns

Schematic of ASTER and the resulting illumation patterns.

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).

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.

Multicolor Imaging Using Far-Red Spectral Demixing

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

Spectral Demixing: Multicolor Imaging with One Laser, One Buffer, and One Acquisition

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 buffers, and the choice of single-molecule-compatible dyes.

To solve this challenge, Abbelight has implemented spectral demixing for SMLM. By separating far-red dyes using a dichroic cube and ratiometric algorithms, spectral demixing elegantly enables simultaneous multicolor imaging in SMLM.


1: A. Mau, K. Friedl, C. Leterrier, N. Bourg, and S. Lévêque-Fort. Fast scanned widefield scheme provides tunable and uniform illumination for optimized SMLM on large fields of view. Nature Communication. May 21, 2020.

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