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

Building 1160, room 424

Objective Mag. / NA Medium
Standard, Olympus UPLXAPO100        100x/1.45, WD 0.13 mm Oil
Lasers Type and Power Selected dyes
405 nm Diode, 1 W DAPI, Hoechst 33342
488 nm Diode, 1 W Alexa488, Atto488, CF488A
561 nm DPSS, 500 mW Alexa 555, dylight 555, CF568
640 nm Diode, 1 W Alexa 647

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)

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