THE COMPANYFemtonics is one of the most dynamically expanding manufacturers of two-photon laser scanning microscopy. We make unique, custom designed 2D systems and as a pioneer we have introduced real-time 3D imaging technology to the market. By our modularity, each Femtonics microscope fits the researcher’s own needs and it can suit a wide variety of biological applications. Our other advantage is our multidisciplinary team which continually enhance and confirm the scientific applicability of our new developments.
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This paper published in Nature Methods demonstrates the capabilities of the Femto3D-AO microscope developed by Gergely Katona et al. The acousto-optic two photon microscope is equipped with an electrically tunable lens focusing the excitation point very fast to any locations in 3D under the objective without mechanical restrictions. Scanning can be done in a near-cubic-millimeter range with high spatial and sub-millisecond temporal resolution. The paper describes two scanning strategies implemented on the system: continuous 3D trajectory scanning allows to precisely follow neural processes over several hundred micrometers to image the propagation of action potentials or dendritic spikes. Random access scanning mode supports imaging evoked or spontaneous calcium-activity in hundreds of neurons simultaneously in vivo.
Roska’s lab (FMI, Basel) mapped the presynaptic network of a V1 orientation selective neuron by using a single-cell-iniciated, monosynaptically restricted, retrograde transsynaptic network tracing with rabies viruses. The labeled neurons expressed GCaMP6s, which allowed to image, in vivo, the visual motion-evoked activity of individual layer 2/3 pyramidal neurons and their whole presynaptic networks across layers in the primary visual cortex of the mice. For the measurement of this large number of neurons scattered across the width of the cortex they used a Femto3D-AO microscope. They found that the neurons within each layer exhibited similar motion direction preferences, forming layer-specific functional modules. In one third of the networks, the layer modules were locked to the direction preference of the postsynaptic neuron, whereas for other networks the direction preference varied by layer.
Marko A. Popovic et al showed new results about electrical behavior of dendritic spines in Nature Communications. The goal of their research was to generate and measure the spread of input signals from the head of an individual spine to the dendrite and in parallel to reveal the role and influence of the neck of the spine. To activate synapses they applied glutamate iontophoresis and two-photon glutamate uncaging. The uncaging experiments were performed with the commonly used MNI glutamate and a new compound, DNI-Glu (produced by Femtonics) which is around 7 times more efficient than the former, even in lower concentrations. The evoked electrical signals (EPSPs) were monitored by electrochromic voltage-sensitive dye simultaneously in the spine and the dendrite. The study provides direct evidence that the resistance of the spine neck is too low to electrically isolate synapses on spine heads, so the synapses of the head electrically behave as if they were located directly on the dendrites.