Thorlabs Bergamo 2-Photon

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General information

  Bergamo 2-photon microscope based on an upright Thorlabs microscope with automatic XY stage, motorized focus and motorized laser wavelength/intensity tuning.

Application
  • 2-channel simutaneous fluorescence imaging of fixed/live samples
  • Multi-channel sequential imaging with tuned excitation
  • CMOS camera for single photon sample inspection
  • High NA 20x water objective to maximum photon collection and spatial resolution
Accessibility
Microscope details
  • Microscope frame: Bergamo II
  • Objectives: Olympus XLUMPLFLN 20X/1.0 W (other objectives are available upon request)
  • Multipoton laser: Chameleon Vision II
  • Excitation tuning range: 680nm - 1080 nm
  • Laser pulse Group Velocity compensition to enhance excitation efficiency
  • Laser intensity control
    • Motorized Laser ND Attenuator: BCM-PA (coarse)
    • Pockel cell: BCM-PCA026 (fine)
  • Detection Capacity: 2 channels from any combinations among 3 detectors
  • Options of combining reflection and transmission signals into single enhanced output
  • Detectors: 3 non-cooled GaAsP detectors
    • Green and red channels in reflection
    • Green channel in transmission
  • Light source for sample inspection: Coolled pE-300 white
  • Brightfield imaging mode: BF
  • Motorised XY Stage:  Thorlabs PLS-XY
  • Focus control
    • Stepper motor focus control (sample focus)
    • 450 um piezo objective scanner (PFM450E, axial scan)
  • Scanner: Gavol-Gavol
  • Auto-focusing: N/A
  • Live experiment support: dark chamber
  • Camera: Thorlabs CS2100M-USB (CMOS: 1920X1080, 16-bit, 50 fr/s)
  • Sample holder: options of slide and petridish
  • Optical bench: active
  • Commercial dervice contract
    • 2-photon laser: Coherent Bronz -July 2022
    • Microscope: pay for engineer visit and broken parts
Filter settings
2-photon setup
Reflection Detector 525/50 607/70
Dichroic in reflection arm 560nm SP
Transmission Detector 525/50 607/70
Dichroic in transmission arm 560 SP

Sample location/imaging setup
Fluorophores DAPI GFP Cy3 Cy5 BF
LED 387 460 555 N/A White LED
Filter Cube Ex: 387/478/555,
BS: 410/504/582/669,
Em: 440/521/607/700
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Workstation and software
  • 2-photon and 1-photon (camera) acquisition software: ThorImage4.0
  • Signal monitoring software (oscilloscope): ThorSync
  • Single photon acquisition software: ThorCam
  • Acquisition workstation
    • CPU: Intel Xeon E5-2620 V3 @2.4GHz
    • GPU: N/A
    • OS: 64-bit Windows 10
    • RAM: 32GB
    • Local data drive (D:): 1 TB
    • Network name: ls-2photon-3d5.ad.susx.ac.uk
Images taken from Bergamo
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