RT Journal Article
SR Electronic
T1 Selective volumetric excitation and imaging for single molecule localization microscopy in multicellular systems
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.12.02.518828
DO 10.1101/2022.12.02.518828
A1 Tommaso Galgani
A1 Yasmina Fedala
A1 Romeo Zapata
A1 Laura Caccianini
A1 Virgile Viasnoff
A1 Jean-Baptiste Sibarita
A1 Rémi Galland
A1 Maxime Dahan
A1 Bassam Hajj
YR 2022
UL http://biorxiv.org/content/early/2022/12/03/2022.12.02.518828.abstract
AB Light sheet fluorescence microscopy (LSFM) has become a leading standard in high-resolution imaging of living samples in 2- and 3-dimensions. Biological samples are however not restricted to a single observation plane and several molecular processes evolve rapidly in 3D. The conventional mechanical scanning required in LSFM limits the range of observable dynamics and are usually restricted in resolution. Here we introduce a new strategy for instantaneous volumetric excitation and volumetric imaging of single-molecules in cell aggregates. The technique combines, for the first time, the use of light sheet microscopy and multifocus microscopy (MFM) and enables a volumetric 4D imaging of biological samples with single-molecule resolution. We engineered the excitation beam to yield a modular and uniform excitation matching the observable detection range of MFM. The strength of the method is highlighted with examples of single-molecule 3D tracking and 3D super-resolution imaging in multicellular samples.Competing Interest StatementThe authors have declared no competing interest.