The control of cellular function and metabolic signalling in astrocytes is complex and highly regulated. Measurement of the nanoscale localisation of protein complexes may be achieved in microscopy by the detection of Förster resonant energy transfer (FRET) between protein-conjugated fluorophores. The use of ‘ensemble’ FRET/FLIM techniques to probe protein-protein interactions in intact cells using functional FRET biosensors such as PercivalHR for metabolic imaging of ATP/ADP. Here we propose the development of a novel microscope system which will enable super-resolved functional FRET measurements within the mitochondria of astrocytes using a combination of high-speed time-correlated single photon counting and single molecule localisation microscopy.
Fili, N., Hari-Gupta, Y., dos Santos, Ália, Cook, A., Poland, S. P., Ameer-Beg, S. M. Parsons, M., Toseland, C. P. (2017) NDP52 activates nuclear myosin VI to enhance RNA polymerase II transcription. Nature Communications., 8(1), p1871.
Poland, S.P., Krstajić, N., Monypenny, J., Coelho, S., Tyndall, D., Walker, R.J., Devauges, V., Richardson, J., Dutton, N., Barber, P.R., Li, D.D-U., Suhling, K., Ng, T., Henderson, R.K., Ameer-Beg, S.M., (2015) A High Speed Multifocal Multiphoton Fluorescence Lifetime Imaging Microscope For Live-cell FRET Imaging, Biomedical Optics Express, 6(2), pp. 277-296.
Simon P. Poland, Grace K. Chan, James A. Levitt, Nikola Krstajić, Ahmet T. Erdogan, Robert K. Henderson, Maddy Parsons, and Simon M. Ameer-Beg, “Multifocal multiphoton volumetric imaging approach for high-speed time-resolved Förster resonance energy transfer imaging in vivo,” Opt. Lett. 43, 6057-6060 (2018)
Bergholt, M. S., Serio, A., McKenzie, J. S., Boyd, A., Soares, R. F., Tillner, J., Chiappini, C., Wu, V., Dannhorn, A., Takats, Z., Williams, A. & Stevens, M. M., Correlated Heterospectral Lipidomics for Biomolecular Profiling of Remyelination in Multiple Sclerosis (2018), ACS Central Science. 4, 1, p. 39-51
Concise Review: The Cellular Conspiracy of Amyotrophic Lateral Sclerosis
Serio, A. & Patani, R., Mar 2018, In : Stem cells (Dayton, Ohio). 36, 6, p. 293-303