Confocal LSM 880

Introduction

The Zeiss LSM 880 with Airyscan offers sensitivity, linear scanning and super-resolution (x1.7 improvement) imaging.

The system features a cooled GaAsP detector and two detectors that complement the spectral working range enabling flexible imaging strategies, including molecular dynamics, recording fluorescence at single photon signal levels and sequential spectral acquisition followed by unmixing of overlapping emission signals.

With the Airyscan detector the system achieves resolution enhancement by use of a radial array of 32 GaAsp detectors which utilizes light that otherwise would be rejected by the confocal pinhole, increasing signal to noise ratio by x4 to x8 times which is utilized to further speed up scanning, get higher sensitivity or increase resolution (by a factor of x1.7) in all 3 dimensions.

In Fast Airyscan mode, four pixel columns are simultaneously imaged utilizing 16 detector elements of the Airyscan detector’s center resulting in x4 faster acquisition with x4 improvement in SNR and x1.5 improvement in resolution.

The LSM 880 is attached to an upright fixed stage microscope (Axio Examiner Z1). The stage carrier can achieve total clearance of 80mm for experimentation on small animals.

Hardware

Lasers

Diode laser	405nm,
Argon multiline	458, 488, 514nm
DPSS laser	561nm
HeNe laser	633nm

Objectives

	Magnification	NA	coverslip	Working distance	Immersion	Parfocal length
Plan Apochromat	x10	0.45	0.17 mm	2 mm	Air	45 mm
Plan Apochromat	x20	0.8	0.17 mm	0.55 mm	Air	45 mm
W Plan Apochromat	x20	1.0	–	1.8 mm	Water dipping	75 mm
W Plan Apochromat	x40	1.0	–	2.5 mm	Water dipping	45 mm
Plan Apochromat	x63	1.4	0.17	0.19 mm	Oil	45 mm

Acquisition

Acquisition is performed with the Zeiss proprietary ZEN system software.

Analysis

The Zen software provides a wide range of 2D/3D image processing tools: export to TIFF, maximum projection, 3D reconstruction, 2D colocalization, spectral unmixing and more. For basic visualization and processing, you can download the free ZEN lite edition software. Offline full versions are available for analysis at CT Computer and Computer IMARIS software. Output files can be opened by Imaris and ImageJ/FIJI for analysis.