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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

 

Specifications

Microscope body

 Eclipse Ti2-E

Stage

 Automated

Perfect Focus system (PFS)

 Yes

Incubator

 Okolab Stage top (25–50⁰C, CO2, humidity including objective heater and temperature sensor)

FOV

 25mm (D)

Objectives

Correction, Magnification, Numerical Aperture, Working Distance

 Plan Apo ʎ 4x NA 0.20 air

 Plan Apo ʎ  10x NA 0.45 air WD 4000µm

 Plan Apo ʎ  20x NA 0.75 air WD 1000µm

 Plan Apo ʎ  40x NA 0.95 air

 Plan Fluor 40x NA 1.30 oil WD 240µm

 Plan Apo ʎ  60x NA 1.40 oil WD 130µm

 Plan Apo ʎ  100x NA 1.45 oil WD 130µm

 10x, 20x, 40x, 60x and 100x magnifications are compatible with DIC and Brightfield

Software

 NIS-Elements software (version 5.42.03 High content analysis package)

RGB Imaging

 Diascopic LED

Fluorescence Imaging

 LED: 395/25nm, 440/20nm, 470/24nm, 510/25nm, 550/15nm, 640/30nm (Lumencor SpectraX)

Detectors

 DS-Ri2 Camera/DS-Qi2 CMOS

 16.25megapixels (not interpolated)

 4908 x 3264 pixel (full-pixel) images at 6 fps, or

 1636 x 1088 pixel (3 x 3 pixel averaging) images at 45 fps

 Imaging format 36mm x23.9mm/6.91 x 4.92mm

Emission filters

 DAPI 460/25nm

 FITC 535/40nm

 Cy5 635/53nm

 Cy7 700/75nm

 Triple emission filer (CFP/YFP/mCherry, Semrock)

 Quad emission filter (LED-DA/FI/TR/Cy5-B, Semrock) 432/36nm, 415/31nm, 596/31nm, 730/139nm

Location

Leichtag building, Room 470