Multi-foci parallelised RESOLFT nanoscopy in an extended field-of-view.

Casas Moreno X, Pennacchietti F, Minet G, Damenti M, Ollech D, Barabas F, Testa I

J Microsc - (-) - [2022-11-14; online 2022-11-14]

Live-cell imaging of biological structures at high resolution poses challenges in the microscope throughput regarding area and speed. For this reason, different parallelisation strategies have been implemented in coordinate- and stochastic-targeted switching super-resolution microscopy techniques. In this line, the molecular nanoscale live imaging with sectioning ability (MoNaLISA), based on reversible saturable optical fluorescence transitions (RESOLFT), offers resolution of large fields of view in a few seconds. In MoNaLISA, engineered light patterns strategically confine the fluorescence to sub-diffracted volumes in a large area and provide optical sectioning, thus enabling volumetric imaging at high speeds. The optical setup presented in this paper extends the degree of parallelisation of the MoNaLISA microscope by more than four times, reaching a field-of-view of 45 - 65 nm$45 - 65\;{\rm{nm}}$ . We set up the periodicity and the optical scheme of the illumination patterns to be power-efficient and homogeneous. In a single recording, this new configuration enables super-resolution imaging of an extended population of the post-synaptic density protein Homer1c in living hippocampal neurons. ( 100 - 130 μ m)2${( {100 - 130\;{\rm{\mu m}}} )^2}$

Ilaria Testa

SciLifeLab Fellow

PubMed 36377300

DOI 10.1111/jmi.13157

Crossref 10.1111/jmi.13157

Publications 9.5.0