Microscope

Upright microscope stand with binocular, motorized X-Y platform for translating microscope stand over a fixed sample (with 35 × 35 mm scanning range), motorized scanning table (with 100 × 50 mm scanning range, image acquisition for ROIs, TileScan, Mark&Find), motorised Z-focus, remote control of X, Y and Z axis of both the stage and stand, piezo drive in Z axis (z-range: 400 µm), filters for epi-fluorescence and brightfield. Bridge platform with manual setting of the focal plane in steps of 6 mm, minimal position 155 mm and maximal position of 235 mm. Interchangeable internal insert for animals, support glass and Petri dish.

 

Available objective:

Nikon CFI75 25×/1.1 WD = 2 mm CC – MP1300

 

Scanning head

Two sets of independent X-Y galvanometric mirrors. Resonant scanner (8 kHz resonant galvanometer). Scanning speed of 30 fps at 512 × 512. A set of X-Y galvanometers for simultanous imaging and uncaging with 1040 nm wavelength.

 

 Illumination path

• Infrared tunable/fixed pulsed laser ChameleonDiscovery TPC (Coherent Inc., CA) with implemented acusto-optic modulator (AOM). Two laser outputs are available:
  • Tunable output has a range of 680 - 1300 nm (step 1 nm), maximum output power 3.5 W, pulsing frequency 80 MHz (± 5), pulse width 100 - 120 fs and with laser power and intensity controlled by AOM.
  • Fixed output has a wavelength of 1040 nm, maximum output power 3.5 W, pulsing frequency 80 MHz (± 5), pulse width 140 fs and with laser power and intensity controlled by AOM.

• Brightfield and fluorescence lamp

 

Detection path

• Reflection Unit
  • Two-channel detection module with optical cables, possibility to set gain and offset for each detector independently, as well as range of each detector.
  • 2 highly sensitive gated GaAsP detectors with quantum efficiency (QE) of over 45 % at 550 nm, detection range of 400-750 nm with low noise and high dynamic range, gating capabilities.

• Transmission Unit
  • 1 transmitted 2-photon light detector suitable for image acquisition simultaneously with other channels
  • 1 GaAsP detector - highly sensitive detector with maximum QE 40 % at 500 nm with quantum efficiency (QE) of over 40 % at 550 nm, detection range of 400-750 nm with low noise and high dynamic range

 

Software

• Prairie View for microscope stand control as well as automatized image acquisition (including single images, time series, z-series, time series, mark and find, tilescans, photostimulation and their combinations), possibility of simultaneous imaging and uncaging. Maximum digital acquisition format 2048 × 2048 pixels.
  • Photostimulation in 2D mode as line scan, segment scan, spiral scan or free hand scan
  • Image acquisition with intensity compensation by laser power or detector voltage
  • A possibility to access data using external software such as Matlab
  • Software is capable of FLIM/PLIM acquisition
  • Spectral scans to detect response of the sample to different wavelengths

• Coherent Software to control the status of the two-photon laser, laser can be controlled with both Coherent Software and Prairie View.

 

HW modules and expansions

• possibility to add external devices to TTLs (for example to do electrophysiology)