Imaging of somata as ROIs in 2D

FemtoSmart Galvo with flexible ROI scanning possibilities, TravellingSalesman

The FemtoSmart Galvo, with its flexible scanning patterns such as 2D random-access point, 2D multiple-line, and folded-frame scanning, supports the manual selection of individual cells in a 2D plane. By skipping measurement of the entire field, it is possible to maintain a high signal-to-noise ratio. Using the TravellingSalesman software module, it is possible to determine the shortest pathway visiting defined points arbitrarily dispersed on the field of view. The short round-trip time results in a high measurement repetition speed, up to 100 Hz for about 30 cells.

Activity measurements on cell
bodies by multiple-line scanning

Activity data along the lines

Activity data as Ca2+ transients

Fast scanning of the entire FOV or an entire volume

FemtoSmart Resonant equipped with Piezo objective positioner kit

Fast-frame scanning based on a resonant scanner combined with fast Z-focusing performed by a Piezo objective positioner kit, is a well-known approach for studying three-dimensional neural networks. In this case, the entire field of view is imaged continuously by the fast scanner, while the objective positioner moves between planes. Different (cortical) layers might be recorded simultaneously this way, or the frames may be assembled to volumes resulting in a four-dimensional dataset.

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Femto3D AcoustoOptic using Anti-motion technology enables the scientist to reveal 3D neural networks in the brain of awake animal models during behavior.

The imaging of cell populations in 3D

Femto3D AcoustoOptic

The Femto3D AcoustoOptic microscopes enable the spatial and temporal complexity of neuronal coding to be resolved by scanning distributed points as regions of interest (ROIs) in a large 3D volume. These microscopes contain the fastest scanner available.

The fastest with Femto3D Atlas

3D random access point scanning, Cell3DFinder

Several thousand cells can be measured near-simultaneously by 3D random-access point scanning by restricting the imaging to sub-regions of the 3D volume. The Cell3DFinder software module finds cell centers in 3D image stacks automatically and can display them as a set of points. The figure (A-D) shows the changes in Ca2+ concentration of 2000 cells from the visual cortex of a GCaMP-expressing mouse as a function of time. See more Wertz et al., Science, 2015; Katona et al., Nat Meth, 2012.

Imaging of multiple somata distributed in 3D in behaving animals

3D chessboard scanning

Chessboard scanning is a planar extension of random-access point scanning using the Anti-motion technology, where random-access points are extended to small squares by drifting the laser beam. These squares can be located anywhere in a near cubic millimeter volume, and include the somata with the surrounding area. Up to 300 somata can be measured simultaneously. The chessboard arrangement of the squares helps to visualize somata, analyze their activity, and correct for motion artefacts. The figure shows in vivo neuronal activity, Ca2+ transients from 100 neuronal somata from the mouse V1 region, labeled with GCaMP6. See also Szalay et al., Neuron, 2016.

Imaging of multiple somata during large motions

3D multi-cube scanning

Multi-cube scanning is a spatially extended mode of chessboard scanning, where a Z dimension is added to the aforementioned squares to cover the whole extent of the somata, therefore preserving all fluorescent information even during large amplitude movements.