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REALM: AO-based localization microscopy deep in complex tissue

Siemons, M. E.; Hanemaaijer, N. A. K.; Kole, M. H. P.; Kapitein, L. C.

2020-06-12 cell biology
10.1101/2020.06.12.147884 bioRxiv
Show abstract

Performing Single-Molecule Localization Microscopy (SMLM) in complex biological tissues, where sample-induced aberrations hamper detection and localization, has remained a challenge. Here we establish REALM (Robust and Effective Adaptive Optics in Localization Microscopy), which corrects aberrations of [≤]1 rad RMS using 297 frames of blinking molecules to improve single-molecule localization. We demonstrate this method by resolving the periodic cytoskeleton of the axon initial segment at 50 m depth in brain tissue.

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