The Opera Phenix can operate in standard fluorescence and spinning disk confocal mode and has temperature and CO2 control for live cell imaging.  It is equipped with 405, 488, 561 and 640 nm solid state lasers and a variety of objectives, including objectives with a high numerical aperture (NA), long working distance (WD) and large field of view (FOV):

• 5x air objective (NA=0.16, WD=12.1 mm, FOV=2583x2583 um) 
• 10x air objective (NA=0.3, WD=5.2 mm, FOV=1292x1292 um)
• 20x air objective (NA=0.4, WD=8.28 mm, FOV=646x646 um)
• 20x water-immersion objective (NA=1.0, WD=1.7 mm, FOV=646x646 um)
• 40x water-immersion objective (NA=1.1, WD=0.62 mm, FOV=323x323 um)
• 63x water-immersion objective (NA=1.15, WD=0.6 mm, FOV=205x205 um)

Microplates

A wide variety of microplates can be used on the Opera Phenix (see also protocols below). For high-resolution imaging, the microplate's bottom of the well should ideally be thin and low. A thin bottom accommodates the short working distance of the water-immersion objectives and limits chromatic and spherical aberration.  A low bottom makes it possible to image wells on the edge of the plate with the water-immersion objectives.  For example, the following microplate is well-suited for high-resolution imaging on the Opera Phenix:

• CellCarrier-96 Ultra Microplates (PerkinElmer Cat # 6055302): bottom thickness = 188 um, bottom height = 200 um.

The following plates are used in CAPB for 3D microtissues:

• 96 well Screenstar microplate (Greiner Bio-One Cat # 655866). Cycloolefin film bottom, thickness = 190 um
• 96 well Sensoplate Plus microplate (Greiner Bio-One Cat # 655891).  Glass-bottom, thickness = 175 um

The microscope's software is used for instrument control (Harmony Instrument Control Software) and for data navigation and analysis (Harmony Office).  All users are encouraged to join user group meetings to develop more efficient protocols for automated image analysis.  To add your name to the mailing list, please contact Jacqueline Escolastico.

Location

Laboratories for Molecular Medicine, Room 528, 70 Ship St, Providence, Rhode Island

Contact information

• Jackie Escolastico (Jacqueline_Escolastico@brown.edu) is the primary contact for training & application support.  To start using the Opera Phenix, please schedule a training session with Jackie.  Jackie can also provide additional information on the associated cell culture facilities and operating procedures. 
• Robbert Creton (Robbert_Creton@brown.edu, webpage) is the Director of the Leduc Bioimaging Facility and Molecular Pathology Core, which will provide operational support for the Opera Phenix.  Please contact Robbert if you have any questions about these centrally available resources or if you need help with automated image analysis using ImageJ.   

Please visit the Leduc Bioimaging Facility website for user rates.

View user fees

For publications that made use of the Opera Phenix, please acknowledge Center funding:  The Opera Phenix in the Center for Animal Alternatives in Testing is supported by the National Institute of Health (U01 ES028184), the National Science Foundation (EPSCoR award 1655221) and a generous gift by Donna McGraw Weiss (’89) and Jason Weiss.  

Trained users can reserve time on the online calendar below. Your training status needs to be updated. This is done on a rolling basis by Jackie Escolastico Jacqueline_Escolastico@brown.edu. Alternatively, you can email Geoffrey_Williams@brown.edu or Robbert_Creton@brown.edu asking for an update of the training status in iLab, listing the microscopes that you have been trained on in the Leduc Bioimaging Facility. 

CAPB users, who are listed on the CAPB website, can sign up for the Opera Phenix any time.  Likewise, approved NSF/EPSCoR and NIH/BRP users can sign up for the Opera Phenix any time. All other users may only sign up for the Opera Phenix in the 48 hour period prior to imaging.

Opera Phenix Signup Calendar

Protocols, user manuals, application guides, and training videos can be found on the password-protected 'protocol page'.

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