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Cyril: Large Field of View Widefield microscope

Cyril: widefield microscope

Summary

Cyril is a widefield microscope with a large field of view and is equipped for 4 imaging modalities: brightfield, Differential Interference Contrast (DIC), color (RGB), and widefield fluorescence. The biggest advantage of this system is the large field of view; every imaging mode benefits from this. This large field of view shows even multiple cells with subcellular details at 100x magnification! Cyril is excellently suited for fixed and live cells. Complete automation in combination with perfect focus system make this an ideal system to acquire entire slides and 96 well plates in an unattended fashion. This could even be done as time lapse with a duration of hours or even overnight and longer.

Image modalities

  • Brightfield
  • DIC imaging
  • Color imaging
  • Widefield fluorescence imaging 

Applications

  • Histology. The color imaging mode is great for fixed tissue slices stained with e.g. hematoxylin and eosin (HE), Periodic acid-Schiff reaction (PAS), Masson's trichrome, Azan stain, and Giemsa. NIS elements software allows easy white balance calibration. We can create an automated acquisition pipeline (JOBS) to acquire mulitple slides in one run. 
  • Immunofluorescence. There are 6 different LED colors with matching filters available in the widefield fluorescence imaging mode. 
  • Fluorescence live cell imaging. In the widefield fluorescence mode there are 6 different LED excitation colors available which are matched with appropriate filters. The stage top incubation chamber regulates temperature, CO2 and humidity.
  • Organoid imaging. The large field of view is ideal to acquire an entire organoid in one frame. In addition, long working distance objectives are available to image through the entire organoid.
  • DIC and brightfield. DIC and brightfield can be used as standalone image modes or can be combined with fluorescence or color imaging. The stage top incubation chamber is fully DIC compatible. 
large field of view of cyril

Technical description

The Nikon Eclipse Ti2-E with 25 mm (d) large field of view, automated stage, perfect focus system, stage top incubator (Okolab) to control the temperature (25 – 50 ⁰C), CO2 and humidity (including objective heater and temperature sensor). The following Nikon objectives are available for this system: Plan Apo lambda 4x NA 0.20 air, Plan Apo lambda 10x NA 0.45 air, Plan Apo lambda 20x NA 0.75 air, Plan Apo lambda 40x NA 0.95 air, Plan Fluor 40x NA 1.30 oil, Plan Apo lambda 60x NA 1.40 oil, and Plan Apo lambda 100x NA 1.45 oil. All imaging functions are integrated in the NIS-Elements software (High content analysis package). 

For RGB imaging, the sample is illuminated with a diascopic LED (Nikon) and images are acquired with a DS-Ri2 camera (Nikon).

For fluorescence imaging, the sample is excited by the Lumencor SpectraX (6 LED colors: 395/25 nm, 440/20 nm, 470/24 nm, 510/25 nm, 550/15 nm, 640/30 nm). The emission light can be filtered by 4 individual emission filters (447/60 nm, 525/50 nm, 609/54 nm, and 698/70 nm), a quad emission filter (LED-DA/FI/TR/Cy5-B, Semrock) or a triple emission filer (CFP/YFP/mCherry, Semrock). The images are acquired with a DS-Qi2 CMOS camera (Nikon).

For DIC and brightfield, the sample is illuminated with a diascopic LED (Nikon). DIC is compatible with 10x, 20x, 40x, 60x and 100x magnifications. TheDS-Ri2 camera (Nikon) or DS-Qi2 CMOS camera (Nikon) can be used for DIC and brightfield. DIC and brightfield can be combined with the fluorescence imaging mode. The stage top incubator is also DIC compatible. 

Location

Leichtag building, Room 470