Selective volumetric excitation and imaging for single molecule localization microscopy in multicellular systems - Laboratoire Physico-Chimie Curie [Institut Curie] Accéder directement au contenu
Pré-Publication, Document De Travail Année : 2023

Selective volumetric excitation and imaging for single molecule localization microscopy in multicellular systems

Résumé

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 singlemolecules 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 singlemolecule 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 singlemolecule 3D tracking and 3D super-resolution imaging in multicellular samples.
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Dates et versions

hal-04048939 , version 1 (28-03-2023)

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Tommaso Galgani, Yasmina Fedala, Romeo Zapata, Laura Caccianini, Virgile Viasnoff, et al.. Selective volumetric excitation and imaging for single molecule localization microscopy in multicellular systems. 2023. ⟨hal-04048939⟩
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