1. Microscope

Upright microscope stand with binocular, motorized X-Y platform for translating microscope stand over a fixed sample (with 35 x 35 mm scanning range), motorized scanning table (with 100 x 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.

 

2. Available of objectives.

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

 

3. Scanning head

Two sets of independent X-Y galvanometric mirrors. Resonant scanner (8kHz resonant galvanometr). Scanning speed of 30 fps at 512x512. A set of X-Y galvanometers for simultanous imaging and uncaging with 1040 nm wavelength.

 

4. Illumination path

• Infrared tunable/fixed pulsed laser Chameleon Discovery 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

 

5. 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

 

7. 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 2048x2048 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.

 

8. HW modules and expansions

Possibility to add external devices to TTLs (for example to do electrophysiology)