Operetta CLS

Operetta
The Operetta CLS is a multi-modal high-content microscopy system. Designed for high throughput, it can capture both widefield and spinning disk confocal images. Setup a single well in a 96 well plate and simply allow the system to replicate those settings across the entire plate. With an automated water immersion objective you also don’t have to sacrifice resolution for high throughput multipoint positioning.
Microscope body: Inveted
Motorised Stage: Yes
Temp Control: Full Enclosure
CO2: Full Enclosure
Software: Harmony

Common Applications

High-Throughput 

The operetta is able to image up to 384 well per assay in a fully automated way 

Live Cell Imaging 

Many biological processes are dynamic events and as such observing living cells is crucial to unravelling and understanding the mechanisms behind them. This requires special consideration when imaging. Firstly, the environmental conditions must be controllable and stable to ensure the cell remain healthy. Secondly, care needs to be taken that the act of imaging has as little impact as possible on the event being observed. 

Spinning Disk Confocal 

Spinning Disk microscopy allows fast confocal-like optical sectioning. By utilising multiple pinholes in a rotating disk several points can be scanned in parallel enabling rapid acquisitions. Especially useful when trying to observe fast biological processes, such as vesicle transport in live cell imaging or thick optically cleared of small spheroids or organoids. 

Colocalisation 

To Complete 

Imaging of Cleared Samples 

Tissue Clearing is a preparation technique with the aim of reducing the inherently scattering nature of tissue. There are several means by which this is achieved but all enable clearer and deeper penetration into the sample using light microscopy. 

FRET 

FRET (Fluorescence Resonance Energy Transfer) is a technique used largely to investigate molecular interactions, or with specially designed reports it can show if molecules are in an active or inactive state. It works by using two carefully selected fluorophores that when in close enough proximity and suitable orientation energy from the donor fluorophore can be transferred to the accepter. This transfer can be observed by monitoring the acceptors emission or the donor fluorescence lifetime. 

Objective Lenses

Magnification 

Numerical Aperture 

Corrections  

Immersion 

Working Distance  

MISC. 

5x 

0.16 

 

Dry 

12.1 mm 

Corrected for 0.17 mm glass bottom 

20X 

0.4 

 

Dry 

8.28 mm 

Corrected for 0 to 1.5 mm glass bottom 

20X 

0.8 

 

Dry 

0.55 mm 

Corrected for 0.17 mm glass bottom 

20x 

1.0 

 

Water 

1.7 mm 

Corrected for 0.17 mm glass bottom 

40x 

1.1 

 

Water 

0.62 mm 

Corrected for 0.17 mm glass bottom 

Light Sources

Source 

Wavelengths 

Nominal Power 

LED 

365 nm 

 

LED 

405 nm 

 

LED 

440 nm 

 

LED 

475 nm 

 

LED 

510 nm 

 

LED 

550 nm 

 

LED 

630 nm 

 

LED 

660 nm 

 

LED 

Transmitted white light 

 

Filters

Name 

Excitation 

Dichroic 

Emission 

Description 

 

BP 500 nm - 550 nm 

 

BP 355 nm – 385 nm 

 

 

BP 430 nm - 500 nm 

 

BP 390 nm – 420 nm 

 

 

BP 655 nm – 705 nm 

 

BP 435 nm – 460 nm 

 

 

BP 685 – 760 nm 

 

BP 460 nm – 490 nm 

 

 

BP 570 – 620 nm 

 

BP 490 nm – 515 nm  

 

 

BP 655 nm – 760 nm 

 

BP 530 nm – 560 nm 

 

 

BP 570 nm – 650 nm 

 

BP 615 nm – 645 nm 

 

 

 

 

BP 650 nm – 675 nm 

 

Detectors

Name 

filter 

Description 

Hamamatsu Orca Flash 4.0 V2 Kamera 

 

CMOS image sensor for scientific measurement.  

Maximum pixel format: 2048 x 2048  

Pixel size 6.5µm (H) x 6.5µm (V) 

Bit Depth:16 bit  

Sensitivity and speed: 100 fps (2048 x 2048 format) 

Miscellaneous

Inserts and specimens: 

  • Stage inserts accommodating 4 histology glass slides, 6-well to 384-well plates 

  • Plastic bottom for low resolution and glass bottom for high resolution imaging  

  • Automated water immersion objectives 

  • Specimen size for 3D imaging: Diameter of cleared sample (50 to 100 um)  

  • Spinning disc pinhole 50 um 

  • Digital phase contrast cannot be used for 3D imaging 

Note: this equipment is not designed for very long live cell imaging, as it doesn’t maintain humidity.