Alba FLIM™ Specifications

Instrument Features

• Dual channel acquisition
• Separate pinholes for each channel for higher resolution
• Computer-controlled alignment of the confocal pinhole and optics
• Choice between scanning mirrors or piezo-controlled XYZ stage
• Single- and multi-photon excitation
• Powered by Vista, a user-friendly software package for the acquisition of FLIM, FRET, RICS, and FCS
• Fully upgradable to Alba FCS™

Measurements

• Fluorescence Lifetime Images (FLIM)
• Fluorescence Resonance Energy Transfer (FRET) efficiency images
• Fluorescence images
• Raster imaging correlation spectroscopy (RICS)
• Steady-state polarization and ratio imaging
• Time courses (kinetics)

Image Parameters Acquired by Alba FLIM™:

• Image area and resolution
• XY and XZ sections
• Speed
• Step size = 50 nm
• Pixels numbers: user selectable from 20 to 1024

Optical Unit

• Light Sources:
  • Single photon lasers housed in a laser launcher with computer-controlled beam expander, laser intensity and shutter; or,
  • Multi-photon excitation with computer-controlled beam expander, laser intensity and shutter
• Optics:
  • Nikon TE2000 inverted microscope with lens revolver for objectives, bottom and side ports and computer-controlled XY-stage for sample slides and for 384-well plates.
  • Available for Leica, Olympus and Zeiss microscopes
  • Objectives:
    • Air objectives with 20X, 40X, 60X magnification and 1.5-8.1 working distances
    • Oil immersion objectives, 1.4 NA and 60X (standard); other aperture available
    • Water immersion objectives,1.2 NA 60X (standard), with coverslip correction (for 0.15-0.18 coverslip); other apertures available
  • Dichroic Filters:
    • For single-photon excitation: 1-, 2-, 3-band filters
    • For multi-photon excitation
    • Polarizer:
    • Cube beam splitter, wavelength range: 450-1100 nm; extinction ratio: 10,000:1 at +/- 3degrees
• Imaging Stage:
  • XYZ piezo-controlled stage, 100x100x50 µm with 5 nm step resolution.
  • Optional Coarse Microscope Stage:
    • Large distance movement (100x100x10 mm)
    • Manual stage or
    • Stepper motor controlled XYZ stage
    • Both stages accommodate
      • Microwell plates
      • Petri dishes
      • Coverslip
• Light Detectors:
  • Photomultiplier tubes
    • Spectral range: 250 nm - 850 nm
    • Dark counts: less than 200 cp
  • Optional APD detectors for RICS
• Laser Scanner Unit:
  • Galvo-controlled mirrors

Modulation Unit

• The modulation and detection unit includes computer-controlled frequency synthesizers (1 -300 MHz) and an RF amplifier.

Data Acquisition

• Data acquisition board
  • Proprietary, 32 bit PCI bus, two-channel card.
• Sampling rate
  • 200 KHz
• Input/Output triggers
  • For monitoring temperature, pH, laser light source fluctuation and starting measurement.

Software: Vista-FCS and Confocal Imaging Microscopy

• Operating system
  • Windows 2000/XP
• Image format
  • Export to ImageJ, MetaMorph
  • Plots export to png, gif, jpeg, bitmap formats

Instrument

• Computer
  • Intel-type CPU, Windows XP operating system
• Power Requirements
  • Universal power input: 110-240 V, 50/60 Hz, 400 VAC
• Dimensions
  • 885 mm (L) x 600 mm (W) x 330 mm (H)
• Weight
  • 40 kg

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