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Sterling: X1 Spinning Disk, TIRF, STORM microscope

Sterling microscope


Sterling is a multifunctional microscope and is equipped for 5 image modalities: spinning disk confocal imaging, Total Internal Reflection Fluorescence (TIRF) imaging, Stochastic Optical Reconstruction Microscopy (STORM), photo activation and stimulation, and bright field. Live cells that express fluorescent proteins at low levels are easily imaged using the spinning disk confocal mode due to the mild illumination method and a sensitive camera. Complete automation in combination with perfect focus system make this an ideal system to acquire live cell pro-longed time-lapses (< 72 h). Biological processes which occur at or near the membrane are best studied using the TIRF mode. In addition, single molecule studies (e.g. tracking or photo physics) are performed using the TIRF mode due to the high-power lasers. Furthermore, super-resolution images are obtained using the STORM modality. This system is fully equipped with stage top incubator for live cells, nonetheless, this system is also suited for fixed cells and non-biological samples. 

Image modalities

  • Spinning disk confocal imaging
  • TIRF (also HILO / ‘dirty’ TIRF)
  • (3D) STORM
  • Photo activation/stimulation
  • Brightfield


Spinning disk confocal imaging

  • Immunofluorescence. Images are acquired fast compared to a laser scanning confocal and the Piezo Z drive allows fast Z stacking.
  • (Fast) live cell imaging. Spinning disk illumination is very mild for your sample, which is beneficial when imaging fluorescent proteins. The stage top incubation chamber regulates the temperature (30 – 40 ⁰C), CO2 and humidity.
  • Pro-longed time-lapse live cell imaging. The perfect focus system controls the focus very precisely and can be used over a long-time course (< 72 h).
  • Filter FRET / ratiometric FRET. Filter FRET using blue, green, red and far red fluorescent dyes or fluorescent proteins can be performed.

TIRF modality

  • Membrane processes in live cells. Processes that occur 200 µm from the cover slip can be very well studied with TIRF.
  • Single molecule tracking in live cells. This can be done near the membrane in full TIRF or relatively higher using HILO TIRF.
  • Single molecule photo physics. Non-biological samples are also compatible on this microscope.

STORM modality

  • STORM and 3D STORM. These super resolution techniques can be performed on live and fixed biological samples. Our analysis workstations are also equipped with the full N-STORM analysis package.

STORM acquisition on N-STORM

Technical description



Microscope body

 Eclipse Ti2-E



Piezo drive

 Mad City Lab Inc. 200µm

Perfect Focus system (PFS)



 Tokai Hit stage top incubator, and full enclosure (30 – 40 ⁰C), CO2 and humidity (including objective heater and temperature sensor)


Correction, Magnification, Numerical Aperture, Working Distance

 Plan Apo ʎ 10x NA 0.45 air

 Plan Apo ʎ 20x NA 0.75 air WD 1000µm

 S Fluor 40x NA 1.30 oil

 Plan Apo ʎ 60x NA 1.40 oil WD 130µm

 Apo TIRF 60x NA 1.49

 SR HP APO TIRF 100x NA 1.49 WD 120µm


 NIS-Elements software (version 5.21.03 High content analysis package)


 405nm, 488nm, 561nm, 640nm (Agilent laser box)

Yokogawa spinning disk

 CSU-X1-50 µm pinholes

Emission filters

 Quad dichroic (DAPI/FITC/TRITC/CY5-for laser lines 405nm, 477nm, 546nm and 638nm)

 432/36nm, 515/30nm, 595/30nm, 755/189nm

 DAPI 455/50nm

 GFP 525/36nm

 TRITC 600/50nm

 Cy5 705/72 nm


 Prime 95B sCMOS camera (Photometrics)

 95% Quantum Efficiency

 11 µm x 11 µm Pixel Area


 18.8 mm FOV


 Diascopic LED





 405nm, 488nm, 561nm, 640nm (Agilent laser box)

Angle control

 N-STORM illumination

Emission Filter

 450/56nm, 525/47nm, 603/44nm, 730/56nm


 iXon Ultra 897 EMCCD camera (Andor)


 Apo TIRF 60x NA 1.49

 SR HP APO TIRF 100x NA 1.49 WD 120µm





 561nm, 640nm

Magnification lenses

 1x, 2x, 4x or 8x (astigmatic lense)


 SR HP APO TIRF 100x 1.49 NA WD 120µm

Photo activation and stimulation

 SDCM/TIRF compatible (digital mirror device by Polygon)

 Light source: AuraII (Lumencor)

 395/25nm, 470/24nm, 555/28nm, and 640/30nm


Leichtag building, Room 470