Fluorescence

Fluorescence Image Caption

Oxidised nanodiamond uptake observed in MCF-7 cells. Cells were incubated with 1 μg / mL oxidized nanodiamonds for 48 hours before fixation. Cells were co-stained with NucBlue nuclear stain and membrane stain Wheat Germ Agglutinin-Alexa Fluor 488. Nanodiamond scattering signal was detected at 633 nm. B J Woodhams, L Ansel-Bollepalli, J Surmacki, H Knowles, L Maggini, M de Volder, M Atatüre and S E Bohndiek. Nanoscale, 10, 12169-12179 (2018).

The study of oxidative stress of cancer cells in culture is made possible through the advent of a number of redox responsive fluorescent proteins and dyes. We are working in cell culture to study the effects of antioxidant capacity and oxidative stress on carcinogenesis using “smart” fluorescent proteins and dyes that give a reactive and responsive readout to changes in glutathione and hydrogen peroxide respectively, using multi-wavelength fluorescence excitation. Long-term experiments (up to 72 h) with laser confocal microscopy are enabled by imaging chambers that maintain temperature, humidity and atmosphere compatible with cell culture. We also exploit Raman spectroscopy to assess biochemical changes, such as oxidative modifications, under these conditions.

In addition to the studies of fluorescence in cell culture, we are advancing near-infrared endoscopy techniques to allow early detection of cancer based on intrinsic contrast and applied fluorescent contrast agents in the gastrointestinal tract.

Lab Members:
Judith WeberDale Waterhouse, Ben Woodhams, Dr Rajesh ShahapureDr Laura Bollepalli.

Collaborators:
Dr Ari Ercole (Addenbrookes Hospital); Dr Elisa Zanier (IRCCS, Italy); Dr Christian Frezza (MRC Cancer Unit); Prof. Mike Murphy (MRC Mitochondrial Biology Unit); Prof. Bruce Ponder (CRUK CI).