ONI Nanoimager

Key features

  • Resolution down to 20 nm allows single molecules to be observed and tracked with high sensitivity.
  • Rapid and precise stabilization of the microscope focus, with locking technology to minimize Z-drift.
  • Post-acquisition drift correction with nanometer precision
  • Precise control and measurement of temperature, with heating elements to keep the entire microscope unit at a stable, uniform temperature.
  • Ideal for live cell imaging, facilitating single-particle tracking.
  • Single molecule localization microscopy in multiple colors and 2 channels. Up to 1W laser power at the source. 8-12 kW/cm² power density at the sample plane.

Location:

The Skou building 1116, room 424

Specifications

Imaging modalities:

  • Single-molecule imaging
  • 3D localization microscopy
  • Förster resonance energy transfer (FRET) spectroscopy

Achievable resolution:

  • Lateral: better than 20 nm
  • Axial: better than 50 nm

3D imaging technique: Astigmatism

Camera: Latest generation sCMOS 82 % peak QE, 1.6 electrons rms read noise at standard scan

  • Magnification: 1 pixel = 117 nm in the sample plane
  • Acquisition speed: 100 fps full frame; 5 kHz with frame height cropped to 2%

sCMOS camera: Hamamatsu ORCA-Flash4.0 V3 Digital CMOS camera

Time for super-resolution full frame: Seconds to minutes (number of localizations and laser power dependent)

    

Simultaneous imaging channels: 1 or 2

Total number of imaging colors: Up to 4 lasers

Field of view: 100 μm x 100 μm (1 channel) or 100 μm x 50 μm (2 channels)

Other light sources: LED for bright-field imaging, NIR auto-focus laser

Emission Channels:

  • Green (channel 0): band 1: 498-551 nm; band 2: 576-620nm
  • Red (channel 1): 665-705 nm

Illumination modes: Closed-loop, continuous illumination angle adjustment between epi-illumination and total-internal reflection

Sample stage: 18/18/9 mm XYZ travel range, closed-loop piezo stage with 2 nm encoder resolution

Temperature control (live cell imaging): Resistive heating, whole instrument

Focus system: One-shot autofocus; Continuous autofocus

Mechanical stability: <1 μm/K drift; <1 nm vibration amplitude (1 Hz to 500 Hz)

Objective Mag. / NA Medium
Standard, Olympus UPLXAPO100     100x/1,45, WD 0,13mm Oil
Lasers Type and Power Selected dyes
405nm Diode, 1W DAPI, Hoechst 33342
488nm Diode, 1W Alexa488, Atto488, CF488A
561nm DPSS, 500mW Alexa 555, dylight 555, CF568
640nm Diode, 1W Alexa 647

Special features

  • Bench-top device; class 1 laser product
  • Homogeneous illumination
  • <10 nm channel mapping accuracy
  • Sensor array (temperature, humidity, CO2, acceleration)
  • Closed-loop adjustments of laser power density at sample plane

Software features

  • Real-time localization analysis and rendering (sCMOS optimized)
  • Real-time FRET trace analysis
  • Automated 3D sample scanning and acquisition
  • Clustering and co-localization analysis
  • Residual drift correction
  • Scripting interface; OMERO compatibility