FEMTOSmart-Resonant Two-Photon Microscope

The FemtoS-Resonant from the FemtoSmart series is equipped with a resonant based scanner and is the most appropriate choice for imaging of the entire field of view (FOV) with high frame rates. In this microscope Femtonics combines the high speed and high sensitivity imaging of living tissues. Resonant scanner based raster scanning is ~5-fold faster for fast acquisition of the entire FOV compared to galvanometric based scanning.

The resonant scanner consists of a fast oscillating mirror for x-axis deflection and a galvanometer mirror for y-axis sweep. Thanks to the 8kHz oscillating speed of the fast mirror, the microscope is capable of gathering images at 31 frames per second.

Features

  • in vivo deep brain imaging down to 850 µm
  • properties of resonant scanner based imaging:
    • 600 µm × 600 µm FOV (with a 20x obj.)
    • raster scanning with 31 fps at 512×512 pixels and 500 fps at 512×32 pixels
    • 16 000 lines/sec (straight, parallel with x-axis)
    • 3D volume scanning with 3 Hz by Piezo objective positioner
    • dynamic pixel dwell time to avoid image distortion
  • custom-designed optical elements for maximal transmission efficiency
  • non-descanned, ultrasensitive GaAsP PMT
  • minimized optical path length by patented travelling detector system
  • high signal-to-noise ratio
  • simultaneous detection of multiple wavelengths
  • compatibility with extended IR wavelength range
  • C -based control software with analysis and upgrade possibilities
  • dedicated software efficiently handling large datasets
  • nearly unlimited measurement times
  • â��F/F calculation
  • parallel recording and analysis of electrophysiological data
  • CMOS camera

High Speed Imaging in a Wide Field of View

In the FEMTOSmart Resonant microscope, Femtonics combines high-speed and high-sensitivity imaging of living tissues by using a fast resonant scanner. Resonant-scanner-based raster scanning acquires images at a 31 frames per second speed for hours, which is ~5 times faster than galvanometer-based scanning in the entire field of view. The velocity of the resonant scanner is non-linear: the speed is different in the center and at the edges of the frame. In the microscope, Pockels cell limits the scanning range to the portion, where scanning velocity is near linear, avoiding photobleaching/photodamage at the two sides of the image. Scan electronics perform dynamic pixel dwelling for data linearization, and to cancel out image distortion.

Scanning modes of FEMTOSmart Resonant

Time Lapse Imaging or Long Term Imaging

While two-photon excitation allows for in-depth imaging and a fine spatial resolution, the high frame scanning rate of the resonant scanner provides a high temporal resolution. This feature means that the microscope is suitable for measuring rapid events in living cells, neural networks, or other circuits. A high frame scanning rate and unlimited video streaming combined with the possibility of performing automated measurements support long-term studies, such as following learning processes, memory retrieval, associative learning, development of model organisms, etc. The middle figure shows an early stage of development of a zebrafish embryo, whose ontogenesis was followed and recorded for over a day, from a low cell state.

3D Volume Scanning 

Fast XY-scanning, combined with a fast movement in the Z plane, ensures the near real-time measurement of a volume in 3D, which enables us to study activity changes in 3D cellular networks or the morphology of organs. Movement in the Z plane can be performed by a Piezo objective positioner or a Liquid lens objective, and two scanning methods are available: frame by frame scanning and n-frame ramping.

Photostimulation

Cells distributed in one or more layers can be stimulated and imaged with high speed using the Resonant microscope equipped with precisely timed LED light sources. LEDs are available at different wavelengths, exciting ChR2 at 473 nm or NpHR at 561 nm, which makes the microscope suitable for performing optogenetic experiments. The microsecond-scale switching time between stimulation and imaging is achieved by using a Pockels cell and gated detectors.

Datasheet

Applications

This product could be used for the following applications:

  • Deep brain imaging
  • Calcium imaging
  • Time-lapse imaging
  • Long-time measurements
  • 3D volume scanning
  • Photostimulation

Manufacturer

This product is manufactured by Femtonics - Advanced Microscopy.

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