Light microscopy core facility

The Light Microscopy Facility provides the CRI with state-of-the-art light microscopy and develops new imaging modes.

The facility specialises in the following areas: 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, second harmonic, fluorescence life-time imaging (FLIM); live imaging at high resolution; quantitative high throughput image acquisition and analysis (e.g. iCys imaging cytometer).

We are also constantly monitoring new developments in imaging techniques, and are actively engaged in developing new applications for cancer research with new instrumentation design. Our further scientific aim is to obtain cancer-relevant comparative imaging results in collaboration with colleagues within the CRI, with newly developed imaging modalities, such as FLIM, non-linear imaging and Raman microscopy, which is John Harris' expertise. He worked on laser development and new imaging instrumentation at the Centre of Biophotonics, Strathclyde, before joining the CRI.

The LaVision TriMScope system has now been equipped with an additional optical parametric oscillator, which provides fs-pulsed MP-excitation ranging from 690nm to 1600nm. The TriMScope is a very sensitive and rapid multi-photon scanning system, which scans the specimen with 64 beams simultaneously. The TriMScope is used mainly for live applications, which Lorraine Berry has developed with CRI research groups.

The CompuCyte iCys research imaging cytometer has become a very popular research tool in the CRI, providing quantitative high-throughput image analysis. Heather Zecchini works on this with CRI researchers. Current applications include ligand uptake in prostate cancer, apoptosis, death receptor involvement in human smooth muscle cells, tumour vasculature and drug distribution and ligand uptake and DNA damage in cancer cells.

Research and development

Current projects include the following:

  1. Second harmonic imaging based on a scattered signal, e.g. to demonstrate the formation of vessels from endothelial cells as well as the extracellular matrix in tumours, and cell behaviour in collagen matrices.
  2. CARS imaging is being combined with fluorescence imaging in cancer drug up-take studies in collaboration with Sumeet Mahajan at the Cambridge Centre for the Physics of Medicine.
  3. Large-area high-efficiency macro imaging using the prototype Mesolens, developed by Brad Amos (MRC Laboratory for Molecular Biology) is being explored by Stefanie Reichelt with the aim of recording subcellular detail throughout 6mm-long mouse embryos and has already been used (in collaboration with Scott Lyons, CRI) to follow individual bioluminescence-labelled tumour cells.

Light microscopy images of mouse tissues using different imaging techniques (Light microscopy core facility report 2010; figure 1)
Figure 1 to 41.
1. 3D volume measurement of the extracellular matrix collagen-vessel relationship in a mouse tumour. Collagen (SHG, blue), vessels (FITC, green), Lectin (mStrawberry, red) (images: L Berry, samples: W English, Murphy lab). 
2 and 3. Multi-photon images of mouse tissues: Ear (2), Kidney (3) expressing a GFP-nuclear marker and Td-tomato membrane marker (images: L Berry, samples: N Berry, MRC-LMB). 
4.Multi-photon image of mouse epidermis (whole mount), two sebaceous glands associated with the hair follicle (image: J Harris, samples: G Donati, Watt lab).


Facility manager
Stefanie Reichelt is the head of the Light microscopy core facility.