Light Microscopy

We specialise in high resolution, high throughput, fluorescence microscopy, three dimensional (3D) digital imaging, time-lapse and low-light-level imaging.

The facility allows access to state-of-the-art equipment including single and multiphoton laser scanning confocals, spinning disc confocal, lightsheet and wide-field microscopes with deconvolution. Many of the systems incorporate environmental chambers allowing temperature-controlled microscopy of living specimens.
We provide support and user training for these systems, complemented by courses and workshops. In addition, we provide assistance with data analysis. Available image analysis packages include 3D image reconstruction, rendering, animation, morphometry, deconvolution, co-localisation and FRET analysis. We also develop bespoke software to address specific analysis problems such as image segmentation and quantification.

We pursue development of light microscopy techniques and their applications in collaborative projects with several Crick research laboratories. These include:

  • Development of specialised imaging techniques, technology and data analysis 
  • Bespoke application of laser scanning confocal microscopy, low-light-level imaging, motion analysis, live cell imaging, spectral imaging, linear unmixing, dynamic imaging of steady state processes (FRAP, paGFP, FLAP), and sensitised emission FRET assay.

Resources

We aim to keep the facility up to date with the cutting-age imaging techniques. Current equipment includes:

Single point laser scanning confocal imaging systems:

  • LSM 880 (Zeiss)
  • LSM 780s (Zeiss) one with a multiphoton tuneable Chameleon Ultra II laser (Coherent)
  • LSM 710s (Zeiss)
  • LSM 510s (Zeiss)
  • SP5s (Leica)one with a Mai Tai HP Deep Sea multiphoton laser (Spectra-Physics) 
Light microscopy image

Light microscopy image

Multi-point laser scanning confocal imaging systems:

  • Swept Field Confocal system Opterra with photoactivation module and a range of laser lines including multiphoton (Bruker)
  • UltraVIEW spinning disk confocal imaging systems (PerkinElmer)
  • Discovery 1 high content screening system (Molecular Devices) adapted in-house for confocal imaging with a Yokogawa scan head.

Wide field imaging systems:

  • Nikon TE2000 based systems with Andor Ixon or Hamamatsu Orca ER scientific CCD cameras
  • Zeiss Axiovert 135TV based system equipped with a microinjection system (Eppendorf).
  • Olympus IX83 and Olympus IX83 TIRF with Hamamatsu ORCA-Flash 4.0 cameras
  • Deltavision (API) systems with Photometrics CCD and EMCCD cameras
  • VS120 slide scanner with Olympus XM10 B/W and Pike F505 C Colour cameras
  • Dual-view lightsheet microscope

Image processing and statistical analysis support is available using:

  • Huygens (SVI)
  • Volocity (Improvision)
  • Imaris (Bitplane)
  • AQM/ iQ (Kinetic Imaging/ Andor)
  • MetaMorph (Universal Imaging)
  • Zen (Zeiss)
  • LAS AF (Leica)
  • Cellprofiler
  • Fiji ImageJ
  • MATLAB (MathWorks)
  •  Mathematica (Wolfram Research)
  •  Neurolucida
  •  Labview (National Instruments).

Kurt Anderson 

Kurt.anderson@crick.ac.uk
+44 (0) 207 269 3091