Atlas combines an intoxicating blend of high-tech science, engineering, refinement in 3D measurements.
The FEMTOSmart series is the next step in classical two-photon microscopy at Femtonics, fully customizable two-photon microscopes.
for three-photon excitation
The three-photon (3P) microscopy allows noninvasive functional imaging of the deeper tissues with higher axial resolution compared to the two-photon excitation. 3P excitation is performed at longer excitation wavelengths which are scattered less in the biological tissues. (See more at 3P imaging) This allows extending the penetration depth, reduces out-of-focus excitation, and increases the SNR.
The 3P range microscopy kit contains:
for Fluorescence Lifetime Imaging
Time correlated single photon counting and the derived fluorescence lifetime imaging measures the time delay between each emitted photon and the laser pulse eliciting it. This time is not affected by fluorophore concentration and excitation intensity fluctuations, however provides intimate information about molecular interactions and dynamics. See more: Applications – FLIM.
Fluorescence lifetime imaging reveals regulation of presynaptic Ca2+ by glutamate uptake and mGluRs, but not somatic voltage in cortical neurons. Olga Tyurikova, Kaiyu Zheng, Elizabeth Nicholson, Yulia Timofeeva, Alexey Semyanov, Kirill Volynski, Dmitri A. Rusakov, Journal of Neurochemistry (2020)
Local Resting Ca2+ Controls the Scale of Astroglial Ca2+ Signals. Claire M. King, Kirsten Bohmbach, Daniel Minge, Andrea Delekate, Kaiyu Zheng, James Reynolds, Cordula Rakers, Andre Zeug, Gabor C. Petzold, Dmitri A. Rusakov, Christian Henneberger, Cell Reports (2020)
Multiplex imaging relates quantal glutamate release to presynaptic Ca2+ homeostasis at multiple synapses in situ. Thomas P. Jensen, Kaiyu Zheng, Nicholas Cole, Jonathan S. Marvin, Loren L. Looger, Dmitri A. Rusakov, Nature Communications (2019)
Polymer microchamber arrays for geometry-controlled drug release: a functional study in human cells of neuronal phenotype. Olga Kopach, Kayiu Zheng, Olga A. Sindeeva, Meiyu Gai, Gleb B. Sukhorukov, Dmitri A. Rusakov, Biomaterials Science (2019)
Glutamate Imaging Reveals Multiple Sites of Stochastic Release in the CA3 Giant Mossy Fiber Boutons. Sylvain Rama, Thomas P. Jensen, Dmitri A. Rusakov, Front. Cell. Neurosci. (2019)
A genetically encoded fluorescent sensor for in vivo imaging of GABA. Jonathan S. Marvin, Yoshiteru Shimoda, Vincent Magloire, Marco Leite, Takashi Kawashima, Thomas P. Jensen, Ilya Kolb, Erika L. Knott, Ondrej Novak, Kaspar Podgorski, Nancy J. Leidenheimer, Dmitri A. Rusakov, Misha B. Ahrens, Dimitri M. Kullmann & Loren L. Looger, Nature Methods (2019)
Monitoring intracellular nanomolar calcium using fluorescence lifetime imaging. Kaiyu Zheng, Thomas P Jensen & Dmitri A Rusakov, Nature Protocols (2018)
Monitoring single-synapse glutamate release and presynaptic calcium concentration in organised brain tissue. Jensen TP, Zheng K, Tyurikova O, Reynolds JP, Rusakov DA, Cell Calcium (2017)
Time-Resolved Imaging Reveals Heterogeneous Landscapes of Nanomolar Ca2+ in Neurons and Astroglia. Kaiyu Zheng, Lucie Bard, James P. Reynolds, Claire King, Thomas P. Jensen, Alexander V. Gourine, Dmitri A. Rusakov, Neuron (2015)
for cultured cells or acute brain slices
Imaging acute brain slices or cultured cells and tissues enables the user to study cells in a controlled environment, outside of a living organism. Gradient contrast illumination eases camera guided patch-clamping while transmitted fluorescence detectors enhance signal collection and SNR.
Features:
Benefits:
for fast Z-stack and 3D imaging
A Piezo objective positioner kit enables the microscope to change the focal point by mechanically moving the objective. With this module, the microscope is able to collect signals from different depths fast enough to resolve biological activity in 3D samples. We can equip most of our microscopes with two types of piezo: piezo specialized for large scanning volumes moves the objective in a 400 µm travel range with up to 30 Hz speed, and the faster one moves it in 100 µm volume with up to 100 Hz (the refresh rate depends on the scanning volume and the size of the objective).
for free rotation of the objective
The motorized tilting module rotates the objective, giving a higher level of freedom to reach the sample from different angles. The module also includes a Piezo objective positioner, ensuring additional movement of the objective in the Z direction.
Novel surgical methods utilizing grin lenses or miniature prisms allow optical investigation of deep structures within the brain. Implementing these technologies is greatly enhanced by our motorized tilting objective:
for multicolor full-field illumination
Wavelength-specific full-field excitation is performed by powerful LED light sources built in a rotating unit above the objective. Fluorescent imaging can be achieved with a wide variety of scientific cameras.
for vessel pattern visualization
Green illumination from an LED light source allows high-contrast visualization of blood vessels, taking advantage of that the highly concentrated heme content of the red blood cells is excitable in the spectral region of 300-650 nm. Using green illumination helps to navigate on the surface of any organs under in vivo conditions and position a patch pipette for bulk loading or patch clamping.
for full-field photostimulation
Full-field illumination using a selected LED source allows molecules and cells to be stimulated over the whole FOV homogeneously. Combine this module with gated detectors to achieve millisecond switching between stimulation and imaging.
See more at Optogenetics
for coarse Z movement
The Bridge structure is a lifting apparatus for FemtoSmart which can replace the foot. It provides extreme freedom in positioning of the body and an increased space under the objective.
for uncaging and optogenetics
The optomechanical design of the light path enables us to direct more beams into the microscope, utilizing the same light path. We offer secondary, fine-tuned laser sources for a wide range of biophotonics applications. See more: Optogenetics, Uncaging.
FEMTO3D Atlas works together with our measurement control and analysis software package called MES. MES is designed in MATLAB with day-to-day lab experiences in the field of cellular and network imaging. The latest version offers new scanning features and improved overall usability. Our developer team is collaborating with researchers from various fields to provide you with a software that makes experimental work efficient.
Key features
Measurement control
AO scanning methods
MES supports coordinate selection and measurement control of various 3D pattern scanning technologies
Module packages
MES can be extended with modules offering various special measurement or analysis functions. We offer MES Premium module pack incorporating functions such as automatic cell selection in Z-stack, data capture automation, photostimulation mapping, and two-channel ratio imaging analysis calculations and more. Ask our experts for more details.
Data access
MES stores data in files based on MATLAB format incorporating all metadata. Besides using MES’ built in analysis features, users can freely access content from the MATLAB IDE as we provide reference to the access functions.
Download
MES has now two development branches separately for galvo and AO high speed recording requirements:
Minimum system requirements
Have you heard about Mesmerize?
For the annotation, downstream analysis and curation of calcium imaging data recorded by Femtonics microscopes, we highly recommend the Mesmerize package written by Kushal Kolar from the Chatzigeorgiou Lab.
The FemtoSmart microscope family is driven by the MESc measurement control and data analysis software. MESc is under active development for performing cellular and network imaging measurements supporting more and more features and Femtonics hardware.
Key features
New MESc features
Measurement control
Data analysis
FEMTO-API Application Programming Interface
Programming Femtonics microscopes
The FEMTO-API is a set of clearly defined instructions that can be given to MESc from various software systems like MATLAB or Python. It lets the experimenter perform data analysis with external tools and control measurements with a freely designable protocol.
DOWNLOAD:
Minimum system requirements
for rodents
Head holder stands and head plates fix the rodent’s head in different positions, enabling precise measurements in the brain. Three types of head holders are available with different dimensions. For anesthetized rodents, we offer a heating pad coupled to a holder, or for behaving animals a stand that ensures access to jetballs, treadmills, or other devices. See more at Behavioral Studies.
Offering maximum flexible adjustment for positioning the head, this holder fixes the scull from one side. The head plate can be moved in three directions of the space and also along the bridge. It is offered for difficult to access parts such as eyes or auditory cortex, and anesthetized rodents.
Stabilized head holder is attached to the rodent’s head at two sides. It is ideal for craniotomy or other surgery where the head of the animal model has to be fixed. It can be coupled to the Gramophone.
for behavioral studies
Gramophone is a single-dimension locomotion tracking device. It allows a head restrained mouse to run on the disk and to respond to visual or other stimuli. The speed and the direction of the running are registered by software-controlled manner. It can be used for high-accuracy velocity recording in conjunction with a two-photon microscope, or as a control interface for behavioral training in a virtual linear maze. The behavior response and the two-photon Ca2+ signals can be recorded simultaneously and aligned with each other.
Technology Overview
This dinitro-indoline-masked form of glutamate releases the bioactive glutamate more rapidly than any other commercially available compound. It was developed for high-quantum yield requiring less irradiation for release, so its effective concentration is lower than other caging scaffolds. The caged compound exists as trifluoroacetic acid salted form (DNI-Glu*TFA) ensuring good solubility, stability and low hygroscopicity. DNI-Glu is a compound developed in-house, only available from Femtonics.
See more at Uncaging.
Endocannabinoid Signaling Mediates Local Dendritic Coordination between Excitatory and Inhibitory Synapses.
Hai Yin Hu, Dennis L.H. Kruijssen, Cátia P. Frias, Balázs Rózsa, Casper C. Hoogenraad, Cell Reports (2019)
A compact holographic projector module for high-resolution 3D multi-site two-photon photostimulation.
Mary Ann Go, Max Mueller, Michael Lawrence Castañares, Veronica Egger, Vincent R. Daria, PLOS One (2019)
Coincidence Detection within the Excitable Rat Olfactory Bulb Granule Cell Spines.
S. Sara Aghvami, Max Müller, Babak N. Araabi and Veronica Egger, J. Neurosci. (2019)
Voltage Gated Calcium Channel Activation by Backpropagating Action Potentials Downregulates NMDAR Function
Anne-Kathrin Theis, Balázs Rózsa, Gergely Katona, Dietmar Schmitz and Friedrich W. Johenning, Front. Cell. Neurosci. (2018)
High efficiency two-photon uncaging coupled by the correction of spontaneous hydrolysis.
Dénes Pálfi, Balázs Chiovini, Gergely Szalay, Attila Kaszás, Gergely F. Turi, Gergely Katona, Péter Ábrányi-Balogh, Milán Szőri, Attila Potor, Orsolya Frigyesi, Csilla Lukácsné Haveland, Zoltán Szadai, Miklós Madarász, Anikó Vasanits-Zsigrai, Ibolya Molnár-Perl, Béla Viskolcz, Imre G. Csizmadia, Zoltán Mucsi and Balázs Rózsa, Org. Biomol. Chem. (2018)
Super-Resolution Imaging of the Extracellular Space in Living Brain Tissue
Jan Tønnesen, V.V.G. Krishna Inavalli, U. Valentin Nägerl, Cell (2018)
Imaging membrane potential changes from dendritic spines using computer-generated holography.
Dimitrii Tanese, Ju-Yun Weng, Valeria Zampini, Vincent de-Sars, Marco Canepari, Balazs J. Rozsa, Valentina Emiliani, Dejan Zecevic, Neurophotonics (2017)
Cell-type–specific inhibition of the dendritic plateau potential in striatal spiny projection neurons.
Kai Du, Yu-Wei Wu, Robert Lindroos, Yu Liu, Balázs Rózsa, Gergely Katona, Jun B. Ding, and Jeanette Hellgren Kotaleski, PNAS (2017)
Electrical behaviour of dendritic spines as revealed by voltage imaging.
Marko A. Popovic, Nicholas Carnevale, Balazs Rozsa; Dejan Zecevic, Nature Communications (2015)
Local Postsynaptic Voltage-Gated Sodium Channel Activation in Dendritic Spines of Olfactory Bulb Granule Cells.
Wolfgang G. Bywalez, Dinu Patirniche, Vanessa Rupprecht, Martin Stemmler, Andreas;V.M. Herz, Denes Palfi, Balazs Rozsa, Veronica Egger, Neuron (2015)
Quantitation of various indolinyl caged glutamates as their o-phthalaldehyde derivatives by high performance liquid chromatography coupled with tandem spectroscopic detections: derivatization, stoichiometry and stability studies.
Vasanits-Zsigrai A, Majercsik O, Toth G, Csampai A, Haveland-Lukacs C, Palfi D, Szadai Z, Rozsa B, Molnar-Perl I, J Chromatogr A. (2015)
Dendritic spikes induce ripples in parvalbumin interneurons during hippocampal sharp waves.
B Chiovini, G F Turi, G Katona, A Kaszas, D Palfi, P Maak, G Szalay, M F Szabo, Z Szadai, Sz Kali and B Rozsa, Neuron (2014)
This dinitro-indoline-masked form of glutamate releases the bioactive glutamate more rapidly than any other commercially available compound. It was developed for high-quantum yield requiring less irradiation for release, so its effective concentration is lower than other caging scaffolds. The caged compound exists as trifluoroacetic acid salted form (DNI-Glu*TFA) ensuring good solubility, stability and low hygroscopicity. DNI-Glu is a compound developed in-house, only available from Femtonics.
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All of our knowledge and results of acousto-optic technology have been poured into our new product, the Femto3D Atlas which has become the peak of the evolution of Femtonics acousto-optic microscopes. Atlas combines an intoxicating blend of high-tech science, engineering, refinement in 3D measurements. Using Atlas, researchers are capable of scanning neuronal, dendritic, and other neuropil activities in 3D about one million times faster compared to classical scanning methods with preserved two-photon resolution thanks to a unique acousto-optical scanner and the superior optical design. Combining its unique fast 3D imaging feature with the property that it implements and goes beyond the traditional galvo and resonant scanner-based microscope imaging functions, Atlas provides an all-in-one solution into the scientists’ hand.
The FemtoSmart series is the next step in classical two-photon microscopy at Femtonics, fully customizable two-photon microscopes. Their special feature is the elevated body which can move in X, Y, and Z directions, providing ample room under the objective for optimal positioning of your sample. This feature makes them suitable for model organisms ranging from zebra fish larvae, through mice navigating in virtual reality to even non-human primates. The microscope’s modular nature allows us to assemble the components, and recombine and upgrade the system to perfectly fit the customer’s needs. FemtoSmart product line contains galvo and/or resonant scanner-based systems which can be equipped with a lot of optional modules enabling it to be adapted to a wide range of biological applications, such as optogenetics, uncaging and dendritic imaging.