Confocal LSM 900 Inverted

Introduction

The Zeiss LSM 900 with Airyscan 2 is a confocal instrument with super-resolution (x1.7 improvement) imaging. The system features two cooled GaAsP detectors for emissions ranging from 400 to 700nm, and a transmitted light detector. With the additional Airyscan 2 detector, the system achieves resolution enhancement (120nm lateral, 350nm axial) from light that would otherwise be rejected by the confocal pinhole that remains open, increasing the signal-to-noise ratio by 4 to 8 times. The split signal is deployed to further speed up scanning, get higher sensitivity, or increase resolution in all 3 dimensions. The Multiplex mode for Airyscan 2 adds new detection configurations for tandem pixel acquisition for fast and gentle acquisition on live samples.

LSM Plus is a complementary imaging function in confocal mode that optimizes acquisition settings and performs linear deconvolution to yield higher resolution and improved SNR.

The system stands on an anti-vibration table and has a top-stage incubation system with temperature and CO₂ control and a high-performance control computer.

Hardware

The LSM900 is attached to an inverted motorized stage microscope (Axio Observer 7) with Definite Focus 3 which is a focus finder that eliminates drifting.

Lasers

	Wavelength	Suitable fluorophores
Solid state laser cassette	640 nm	Cy5, APC, Alexa 647, TO-PRO-3
Solid state laser cassette	561 nm	Cy3, TAMRA, PI, Texas Red, PKH26
Solid state laser cassette	488 nm	Cy2, GFP, Mitotracker Green, JC1
Solid state laser cassette	405 nm	DAPI, Hoechst

Objective lenses

Position	Objective lens	Magni-fication	NA	coverslip	Working distance	Immersion	Parfocal length
1	EC Plan Neofluar	x5	0.15	0.17mm	18.5mm	Without	45mm
2	Plan-Apochromat	x10	0.45	0.17mm	20mm	Without	45mm
3	Plan-Apochromat	x20	0.8	0.17mm	0.55mm	Without	45mm
4	LD LCI Plan-Apochromat	x25	0.8	0.17mm	0.57 at cover glass 0.17	Water, Silicone Oil, Glycerine, Oil	45mm
5	Plan-Apochromat	x63	1.4	0.17mm	0.19mm	oil	45mm

Acquisition

Acquisition is performed with the Zeiss proprietary ZEN Blue 3.5 system software.

Analysis

The Zen Blue software provides a wide range of 2D/3D image processing tools: export to TIFF, maximum projection, 3D reconstruction, 2D colocalization, image stitching 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.

New users

Please contact Maya Holdengreber, tel. 073-378-1106, to coordinate a meeting.
Tips for immunofluorescence experiments.
Please prepare controls before starting a new model.

Reservations

For general and live cell confocal imaging including tiling and z sections, the LSM900 Inverted can be reserved through our BookItLab online reservation system.