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 which remains open, increasing 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 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.
| Laser line | 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. An offline full version is available for analysis at Computer CT analysis. Output files can be opened by Imaris and ImageJ/FIJI for analysis.
New users
Please contact Maya Holdengreber, tel. +972-73-3781106, 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 in the BookItLab online reservation system following instruction.
Reading material before first instruction session
Instruction LSM900 inverted short
Confocal laser scanning principles
Airyscan