This project is co supervised with Madeline Lancaster

Fluorescence microscopy provides images of living cells and tissues with molecular specificity, but comes with a terrible price: premature death. A typical experiment may kill the sample within a few hours, or even minutes. Using advanced techniques, such as light sheet illumination with the newest fluorophores, can prolong life up to a few days. However, even the best current approaches cannot image the full, six-month process of neuronal maturation within developing human brain tissue. This project is aimed at realising this goal.

You will investigate the photophysics of light-induced death (phototoxicity) and use your results to find optimal experimental parameters for any experiment using fluorescent labels in living samples. Using the advanced light microscopes available at the LMB, you will use novel fluorescent molecules to image the early stages of neuronal growth. In parallel, you will construct a microscope that uses scattered light to image thick samples with high resolution. This will then be combined with a fluorescence imaging system to provide correlative imaging over the full cycle of neural development within a model brain organoid.

You will work synergistically with two teams: one expert in advanced microscopy development (James Manton) and the other in studying neurodevelopment using cerebral organoids (Madeline Lancaster). By design, the photophysical investigations are broad in scope, allowing results to be applied to a variety of imaging scenarios. As such, we envisage the project expanding beyond neural development to study, with collaborators, other challenging dynamic processes in different model systems. In addition, there is scope for using the long-term organoid imaging platform you will develop to study differences in brain growth between species and in neurodevelopmental disorders, such as microcephaly.

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References

Cranfill PJ, Sell BR, Baird MA, Allen JR, Lavagnino Z, de Gruiter HM, Kremers G, Davidson MW, Ustione A, Piston DW. (2016)
Quantitative assessment of fluorescent proteins
Nature Methods. 13(7):557–562

Scholler J, Groux K, Goureau O, Sahel J, Fink M, Reichman S, Boccara C, Grieve K. (2020)
Dynamic full-field optical coherence tomography: 3D live-imaging of retinal organoids.
Light: Science & Applications. 9(140)

Chang BC*, Manton JD*, Sapoznik E, Pohlkamp T, Terrones TS, Welf ES, Murali VS, Roudot P, Hake K, Whitehead L, York AG, Dean KM, Fiolka R. (2021)
Real-time multi-angle projection imaging of biological dynamics
Nature Methods. 18(7):829-834.

Benito-Kwiecinski S, Giandomenico SL, Sutcliffe M, Riis ES, Freire-Pritchett P, Kelava I, Wunderlich S, Martin U, Wray G, McDole K, Lancaster MA. (2021)
An early cell shape transition drives evolutionary expansion of human forebrain.
Cell. S0092-8674(21)00239-7.

Giandomenico SL, Mierau SB, Gibbons GM, Wenger LMD, Masullo L, Sit T, Sutcliffe M, Boulanger J, Tripodi M, Derivery E, Paulsen O, Lakatos A, Lancaster MA. (2019)
Cerebral organoids at the air-liquid interface generate diverse nerve tracts with functional output.
Nat Neurosci. 22(4): 669-679.

Lancaster MA, Renner M, Martin C-A, Wenzel D, Bicknell LS, Hurles ME, Homfray T, Penninger JM, Jackson AP, Knoblich JA. (2013)
Cerebral organoids model human brain development and microcephaly.
Nature. 501(7467):373-9.