Light microscopy core facility

The light microscopy facility provides training and support to scientists in a wide-range of imaging acquisition and analysis techniques. Instruments in the Light Microscopy Core range from wide-field imaging systems to non-linear multi-photon and lifetime imaging systems for cell and intra-vital tissue studies.

The facility is specialised in the following areas of expertise:

  • Advanced live-cell imaging using wide-field and spinning disc imaging systems
  • High-quality confocal scanning light microscopy
  • Non-linear imaging techniques such as multi-photon and second harmonic imaging
  • Fluorescence life-time imaging (FLIM)
  • Live-animal imaging at high resolution
  • Quantitative high throughput image acquisition and analysis (e.g. iCys)

 

 


Confocal microscopy

The facility has two Nikon and two Leica confocal laser-scanning systems for high-quality fluorescence imaging. The Leica systems have additional fast resonance scanning modes, whilst the Nikon systems have the advanced spectral detection option of imaging 32-channels simultaneously. Three of the confocal systems have large area scans or multiple time-point experiments with CO2 and temperature-controlled incubation chambers.

Wide-field imaging systems

Time-lapse observation systems are available for studies of apoptosis and growth development (>24 hours). These microscopes have CCD or EMCCD cameras, high-quality DIC optic or phase contrast, perfect focus, and multi-position stages. Solent or Tokai-hit portable chambers for CO2 humidity and temperature control and an Eppendorf microinjector are available.

Spinning disc system

Andor DRAGONFLY – Fast 2D – 6D imaging

The Dragonfly offers you a sharp insight into the world of your specimen by employing a novel multi-point confocal system, optimized for fast, flexible and sensitive scanning. It extends your sample’s longevity and significantly speeds up data acquisition.

As well as confocal, the system provides further extraordinary control of illumination by offering enhanced widefield and TIRF microscopy, extending the reach from nm to mm and from single molecules to cells, tissues, organoids and embryos.

Multi-photon microscopy

Multi-photon imaging is the state-of-the-art technique for the observation of living mammalian cells and special physiological probes within tissues.  It is highly recommended for monitoring thick biopsies, as the depth penetration of IR is greater than for shorter visible wavelengths.

The facility has two multi-photon systems: (1) the Leica SP5, equipped with a pulsed Coherent Chameleon Ultra II laser infra-red laser.  (2) The LaVision BioTec ‘TriMscope’, with a 4W Coherent Ultra II (690 – 1080nm) and an additional APE OPO, extending the excitation wavelength from 1050 to 1580nm. This allows for background-free deep tissue imaging of fluorescence ranging from UV to IR-probes and second harmonic imaging e.g. of collagen.

High throughput image acquisition and analysis: Perkin Elmer Operetta CLS

The Operetta CLS system combines speed and sensitivity with the powerful and intuitive data analysis delivers everything you need from scanning a few slides at high-resolution with FISH of RNAi labels all the way to multi-well high-content drug and target analysis.

What’s more, the Operetta CLS system is part of a comprehensive HCS workflow – everything from HCS systems and microplates to automation and informatics for every application. THe Harmony® high-content imaging and analysis software is easy-to-learn, easy-to-use software that empowers biologists to do their own analysis

  • Cell-based screening and phenotyping with user-friendly all-in-one image analysis pipeline
  • High content imaging of live cells and organoids
  • Multi-well plates and slides (1-4 slides or IBIDI/Labtek slides or multi-well plates)
  • RNAscope imaging at high-resolution in large tissue samples or cells
  • IDEAL for highly multiplexed immunofluorescence imaging up to 24 colours
  • Optimized for RNAscope probes with 8 LEDs for 8 colour imaging
  • Flow cytometry-type analysis on a slide with fast and high-resolution options
  • High precision Re-Scans of the same sample (e.g. re-stained) with alignment algorithms
  • Wide-field and CONFOCAL imaging options for 3D volume imaging