Confocal LSM 900

Introduction

The Zeiss LSM 900 with Airyscan 2 is a confocal instrument with super-resolution imaging with x1.7 improvement in all three dimensions.

The system features two cooled GaAsP detectors for emissions ranging from 400 to 700nm and a transmitted light detector. With the Airyscan 2 detector, the system achieves resolution enhancement by use of a radial array of 32 concentrically arranged detection elements which utilizes light that otherwise would be rejected by the confocal pinhole which remains open, increasing the signal to noise ratio by x4 to x8 times (120nm lateral, 350nm axial). The split signal is deployed to further speed up scanning, get higher sensitivity, or increased resolution. The Multiplex mode for Airyscan 2 adds new detection configurations for parallel pixel acquisition for fast and gentle acquisition on live samples.

 

Hardware

The LSM900 is attached to an upright microscope (Axio Examiner Z1). The stage carrier is fixed.

Solid state lasers

Wavelength Suitable fluorophores
640 nm Cy5, APC, Alexa 647, TO-PRO-3
561 nm Cy3, TAMRA, PI, Texas Red, PKH26
488 nm Cy2, GFP, Mitotracker GreenOhter
405 nm DAPI, Hoechst

Objective lenses

Position Objective lens Magni-fication NA coverslip Working distance Immersion Parfocal length
1 W Plan Apochromat x20 1.0 0 1.8 mm Water dipping 75 mm
2 W Plan Apochromat x10 0.5 0 3.7 mm Water dipping 75mm
3 EC Plan Neofluar x5 0.16 0.17mm 18.5mm Without 45mm
4 W Plan Apochromat x40 1.0 0 2.5 mm Water dipping 45 mm

Objectives from one of our other systems can be installed for a particular experiment by appointment.

Acquisition

Acquisition is performed with the Zeiss proprietary ZEN 3.0 system software.

 

Analysis

The Zen Blue 3.0 software provides a wide range of 2D/3D image processing tools: export to TIFF, maximum projection, 3D reconstruction and more. For basic visualization and processing, you can download the free ZEN lite edition software.
An offline full version is available for processing and analysis at Computer CT software. Output files can be opened by Imaris and ImageJ/FIJI for analysis.

 

New users

Please contact Maya Holdengreber, tel. +972-77-8871106, to coordinate a meeting.
Tips for immunofluorescence experiments.
Please prepare controls before starting a new model.

 

Reading material before first instruction session

Instruction LSM900 short
Confocal laser scanning principles
Airyscan