Chu, L.A., Lu, C.H., Yang, S.M., Liu, Y.T., Feng, K.L., Tsai, Y.C., Chang, W.K., Wang, W.C., Chang, S.W., Chen, P., Lee, T.K., Hwu, Y.K., Chiang, A.S., Chen, B.C. (2019). Rapid single-wavelength lightsheet localization microscopy for clarified tissue.  Nat. Commun. 10(1): 4762.

FBrf0243807

Research paper

Optical super-resolution microscopy allows nanoscale imaging of protein molecules in intact biological tissues. However, it is still challenging to perform large volume super-resolution imaging for entire animal organs. Here we develop a single-wavelength Bessel lightsheet method, optimized for refractive-index matching with clarified specimens to overcome the aberrations encountered in imaging thick tissues. Using spontaneous blinking fluorophores to label proteins of interest, we resolve the morphology of most, if not all, dopaminergic neurons in the whole adult brain (3.64 × 107 µm3) of Drosophila melanogaster at the nanometer scale with high imaging speed (436 µm3 per second) for localization. Quantitative single-molecule localization reveals the subcellular distribution of a monoamine transporter protein in the axons of a single, identified serotonergic Dorsal Paired Medial (DPM) neuron. Large datasets are obtained from imaging one brain per day to provide a robust statistical analysis of these imaging data.

PMC6800451 (PMC) (EuropePMC)