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.

Overview

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.

Examples of work
  • Olympus FVMPE-RS microscope
    The Olympus FVMPE-RS at the Institute
  • Multi-coloured tumours indicate cells invading a lung
    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
  • Microscopy image of the thymus
    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
  • Microscopy image of blood vessels in the eye
    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

Video: Captured on the Olympus FVMPE-RS, green and magenta ‘tendrils’ show the network of blood vessels that are essential for the eye to form. Credit: Stephen Mieruszynski and Leigh Coultas

Hardware

Objective magnification 5x 10x 20x 25x 25x
Immersion media Air Multi Water Water Clarity
XY-resolution (µm) 4.97 2.49 1.24 0.99 0.99
Working distance (mm) 20 8 2 2 8
NA 0.10 0.60 1.00 1.05 1.00

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

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.

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

Resources

Nikon: Fundamentals and applications of multiphoton microscopy 

Video: A dissected retina image has been carefully removed to preserve its 3D structure, then imaged using a multiphoton microscope to allow the full structure to be acquired. Previously, imaging a retina would require the sample to be flattened. Credit: Leigh Coultas, Stephen Mieruszynski, Lachlan Whitehead