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Centre for Dynamic Imaging

Zeiss LSM 880 NLO with fast Airyscan Confocal

A multifunctional system suited for studying diseases in their host environment.

The Zeiss LSM 880 NLO with fast Airyscan Confocal microscope is a well-equipped multi-photon system suitable for in vivo and in vitro studies of disease.

The system is equipped with a MaiTai DeepSee multi-photon laser tuneable from 690 nm – 1040 nm. This provides an excellent system well suited to intravital microscopy.

The microscope also features an Airyscan detection unit for an improved resolution of 1.7x that of diffraction limited confocal microscopy.

Equipment booking
Above: To image this whole lymph node in 3D, the organ was rendered optically transparent to permit a better penetration of the light for fluorescence microscopy imaging. Credit: Fanny Lafouresse and Verena Wimmer
Above: Intestinal villi imaged on the Zeiss LSM 880 microscope. Credit: Caleb Dawson

Objectives

Light sources

  • Diode laser – 405 nm
  • Argon laser – 458 nm, 488 nm, 594 nm
  • DPSS – 561 nm
  • HeNe – 633 nm
  • MaiTai DeepSee – 690 nm – 1040 nm

Detection source

  • 2 x Big (GaAsP) NDD detectors
  • 2 x NDD detectors
  • 1 x 32 channel GaAsP array
  • 1 x Airyscan Detector
Objective size20254063
NA10.81.31.4
Immersion mediaWater (dipping)Multi
Water/Glycerine/Oil		OilOil
XY-resolution (μm) (@ 488 nm)0.2970.372/0.372/0.3720.2290.212
Z-resolution (µm) (@ 488 nm)1.32.031/2.246/2.3140.8760.755
Working distance (mm)1.70.5700.2100.190
Above: This image captures various immune cells, identified by membrane staining, interacting together within a lymph node. Credit: Fanny Lafouresse and Verena Wimmer
Above: Cross section of a developing five-day old zebrafish intestine. Single mitochondria can be seen as small green donuts throughout the image. DNA within the nucleus (blue) and actin cytoskeleton (red) enable us to identify single cells. Credit: Stephen Mieruszynski

Technological specifications (capabilities)

Available modalities

  • Z-stack
  • Time-series
  • Tile scan
  • Multi positions
  • FRAP
  • Spectral unmixing
  • Online fingerprinting

Additional hardware

  • Airyscan detection
  • Fast Airyscan module

Unique features

The Zeiss LSM 880 NLO plus fast Airyscan Confocal microscope is a multi-functional two-photon microscope with numerous detection options including the Zeiss Airyscan and fast Airyscan modules for high resolution and high speed imaging respectively.

Above: 3D imaging enables researchers to visualise bone metastasis progression at single-cell resolution. Credit: Raymond Yip
Above: A group of red blood cells (magenta) infected by a malaria-causing parasite. Parasite DNA is shown in blue, some parasite proteins are shown in green. Credit: Michał Pasternak

Limitations

  • Not suited for live cellular samples
  • Photobleaching
Above: A Zeiss LSM 880 microscope at WEHI
Above: High resolution 3D imaging showing tumour cells induce the formation of new blood vessels to facilitate their growth within bone marrow. Credit: Raymond Yip

Related microscopes

Case studies

Experts

Professor Kelly Rogers
  • Head, Centre for Dynamic Imaging
Dr Niall Geoghegan
  • Lattice light sheet specialist
Contact us

Contact the Centre for Dynamic Imaging to book equipment or discuss imaging collaborations.

imaging@wehi.edu.au

Centre for Dynamic Imaging

Olympus FVMPE-RS Multiphoton

The Olympus FVMPE-RS allows both rapid and deep image acquisition, giving scientists great potential for imaging samples that would be impossible to examine on traditional microscopes.

The Olympus FVMPE-RS Multiphoton system is an upright microscope utilising dual pulsed two-photon far-red lasers for deep tissue imaging. The system also includes a resonant scan head allowing for extremely rapid imaging for tracking cell migration in living tissue.

The Olympus system is capable of imaging a wide range of endogenous and antibody labelled samples and generating second harmonics of unstained structures in whole tissue. The system also has special objective lenses designed to image cleared tissue to image up to 8 mm into organs.

The system is compatible with most samples but is ideally suited for preclinical models and large tissue imaging, giving scientists a chance to image deeper and over prolonged periods of time.

Equipment booking

Hardware

Light sources

  • Olympus U-HGLGPS halogen light source + generic DAPI/GFP/RFP filters
  • Mai-Tai eHP DeepSee multi-photon laser
  • Insight DS+ multi-photon laser

Detection sources

  • 2 x PMT detectors
  • 2 x GaAsP detectors
Objective magnification5x10x20x25x25x
Immersion mediaAirMultiWaterWaterClarity
XY-resolution (µm)4.972.491.240.990.99
Working distance (mm)208228
NA0.100.601.001.051.00
Above: The Olympus FVMPE-RS has allowed researchers to study the natural undulations and structure of the thymic epithelial cell network at cellular resolution. Credit: Julie Sheridan, Edwin Hawkins
Above: A hyaloid blood vessel network nourishes the eye during its development. Understanding how these vessels naturally die off once development is complete may shed light on diseases such as diabetic retinopathy and age-related macular edema. Credit: Stephen Mieruszynski, Leigh Coultas

Technological specifications (Capabilities)

Available modalities 

  • Z-stack
  • Time-series
  • Tile scan
  • Multi positions

Additional hardware

  • Resonant scan head
  • SIM scanner

Unique features

  • The system has a structured illumination microscopy (SIM) scanner, allowing for simultaneous imaging with one laser while using the other laser for photo-conversion or cell-ablation of small areas.
  • The MATL (Multi-Area Time Lapse) feature allows for highly customisable experiments of multiple areas.
  • The deep focus mode allows users to choose between depth of imaging and optimal Z-resolution.
Above: The Olympus FVMPE-RS at the Institute
Above: To see how cancer cells travel to new organs, researchers grew multi-coloured tumours that allowed them to track each cell invading a lung. Credit: Caleb Dawson

Limitations

  • Lower resolution in Z-axis than traditional confocal
  • High laser powers can be detrimental to sample integrity
  • Broad excitation profiles with single laser increase possible channel cross-talk
  • Excitation profiles not characterised for all fluorophores

Resource

Nikon: Fundamentals and applications of multiphoton microscopy

Acknowledgement

This microscope was purchased with the generous support of the Australian Cancer Research Foundation (ACRF).

Related microscope

Case study

Experts

Professor Kelly Rogers
  • Head, Centre for Dynamic Imaging
Dr Lachlan Whitehead
  • Bioimage analyst
Dr Niall Geoghegan
  • Lattice light sheet specialist
Contact us

Contact the Centre for Dynamic Imaging to book equipment or discuss imaging collaborations.

imaging@wehi.edu.au

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