Confocal Microscopes: Zeiss LSM 980 NLO Airyscan 2 confocal/two-photon laser-scanning microscope (508D)

The Zeiss LSM 980 NLO Airyscan 2 confocal/two-photon laser-scanning microscope is an inverted laser-scanning system that features the Airyscan 2 super-resolution detector which enables up to 90-nm resolution with Airyscan Joint Deconvolution and includes the SR-4Y and SR-8Y multiplex modes for fast parallel imaging at frame rates of up to 47 fps, a 34-channel spectral array consisting of a 32-channel GaAsP PMT spectral detector and two multi-alkali PMTs, two internal GaAsP PMT detectors in the BiG.2 module for confocal, FCS, and photon-counting experiments, two external non-descanned GaAsP PMT detectors for two-photon microscopy, temperature and CO2 control, six visible laser lines for single-photon excitation, and the tunable Spectra-Physics Mai Tai eHP DeepSee Ti:sapphire laser for two-photon excitation.

Features:

Airyscan 2 super-resolution detector, including SR, SR-4Y, SR-8Y, and CO-8Y modes
34-channel QUASAR spectral PMT array (32-channel GaAsP + 2 x MA-PMTs)
Binary GaAsP module (BiG.2) with 2 x GaAsP PMTs for 500-550 and 570-610 nm detection
2 x external GaAsP PMTs for two-photon detection (500-550 and 570-610 nm)
Transmission PMT detector
Temperature and CO2 control in a dark incubation chamber
Motorised XY and Z stages
Definite Focus.2 for focus stabilisation in long-term imaging

Objectives:

Fluar 2.5x/0.12 M27 air / 8.7 mm WD / 3.315 μm/pixel (1024x1024)
Plan-Apochromat 10x/0.45 M27 air / 2 mm WD / 0.829 μm/px (1024×1024)
Plan-Apochromat 20x/0.8 M27 air / 610 µm WD / 0.415 µm/pixel (1024×1024)
C-Achroplan 32x/0.85 W Corr M27 VIS-IR / 1.1 mm WD /0.259 µm/pixel (1024×1024)
C-Apochromat 40x/1.2 W Corr M27 FCS / 280 µm WD / 0.208 µm/pixel (1024×1024)
C Plan-Apochromat 63x/1.4 Oil DIC M27 / 140µm WD / 0.132 µm/pixel (1024×1024)

Lasers:

405 nm (DAPI/BFP)
445 nm (CFP)
488 nm (GFP / Alexa Fluor 488)
514 nm (YFP)
561 nm (Cy3 / mCherry / Alexa Fluor 546/555/568)
639 nm (Cy5 / Alexa Fluor 647)

Left: GATTA-Cells huFIB specimen showing mitochondria (Tom 20 with Alexa Fluor 488), microtubules (a-Tubulin with Alexa Fluor 555), and actin (Alexa Fluor 647) after LSM Plus processing, which enables up to 160-nm resolution with a pinhole size of ~0.8 Airy unit (AU), and 120-nm resolution with a pinhole size of ~0.3 AU. Middle: The same specimen after Airyscan processing, which enables up to 120-nm resolution in full super-resolution mode (1.25-AU equivalent). Right: Resolution test using GATTA-SIM 90B nanorulers specimen after Airyscan Joint Deconvolution, which enables up to 90-nm resolution.

To cite the use of the Zeiss LSM 980 NLO Airyscan 2 confocal/two-photon laser-scanning microscope in a manuscript, please refer to the text below:

“Image acquisition was performed using a Zeiss C Plan-Apochromat 63x/1.4 NA oil-immersion objective on a confocal/two-photon laser-scanning microscope (LSM 980 NLO Airyscan 2, Carl Zeiss GmbH, Germany) built around an Axio Observer 7 body and equipped with an Airyscan 2 super-resolution detector, a 34-channel spectral PMT (photomultiplier tube) array, two internal GaAsP PMTs, a transmission PMT, and two external GaAsP PMTs for non-descanned detection in two-photon microscopy, and controlled by Zen Blue software.”