Fast two-layer two-photon imaging of neuronal cell populations using an electrically tunable lens
By Benjamin F. Grewe, F. Voigt, Marcel van ’t Hoff, Fritjof Helmchen. 2011
306 citations recorded by the source corpus.
Functional two-photon Ca2+-imaging is a versatile tool to study the dynamics of neuronal populations in brain slices and living animals. However, population imaging is typically restricted to a single two-dimensional image plane. By introducing an electrically tunable lens into the excitation path of a two-photon microscope we were able to realize fast axial focus shifts within 15 ms. The maximum axial scan range was 0.7 mm employing a 40x NA0.8 water immersion objective, plenty for typically required ranges of 0.2–0.3 mm. By combining the axial scanning method with 2D acousto-optic frame scanning and random-access scanning, we measured neuronal population activity of about 40 neurons across two imaging planes separated by 40 μm and achieved scan rates up to 20–30 Hz. The method presented is easily applicable and allows upgrading of existing two-photon microscopes for fast 3D scanning.
Subfields: Biophysics, Cellular and Molecular Neuroscience
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Citations in the corpus
- Re-evaluating circuit mechanisms underlying pattern separation (2019, neutral): “Several promising new imaging technologies that can record intracellular calcium from large numbers of identified cells have also been recently developed (Grewe et al., 2011; Katona et al., 2012; Froudarakis et al., 2014; Nadella et al., 2016; Sofroniew et al., 2016; Yang et al., 2016).”
- Simultaneous multi-plane imaging of neural circuits (2016, unknown): “The standalone ETL, when properly inserted into the microscope, represents perhaps the most cost effective solution for fast focusing.”
- A large field of view two-photon mesoscope with subcellular resolution for in vivo imaging (2016, background): “Electrotunable lenses introduce spherical and chromatic aberrations (Grewe et al. 2011).”
- Fast functional imaging of multiple brain regions in intact zebrafish larvae using Selective Plane Illumination Microscopy (2013, background): “In the last two decades, functional imaging approaches, in which the neurons’ spiking dynamics is monitored via the increase in fluorescence of calcium-binding reporters, have allowed to partially circumvent this limitation (Grienberger and Konnerth, 2012).”