Opera Phenix High-Content Screening System
The Opera Phenix High-Content Screening System is a microscope designed for high-throughput 3D imaging of fluorescent samples in microplates. This imaging system, open to all investigators, is critical to the mission of the Center to Advance Predictive Biology and associated NSF/EPSCoR and NIH/BRP awards.
- Microscope Configuration
- Image Analysis
- Location & Contacts
- Publication & Funding
- Signup Calendar
- Additional Information
Microscope configuration. 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
Image Analysis. The microscope's software is used for instrument control (Harmony Instrument Control Software) and for data navigation and analysis (Harmony Office). In addition, Imaris image analysis software is available for visualization, analysis, segmentation and interpretation of 3D datasets. 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 Susan Hall.
Laboratories for Molecular Medicine, Room 528, 70 Ship St, Providence, Rhode Island
- Samanta Madnick (Samantha_Madnick@brown.edu) is the primary contact for training & application support. To start using the Opera Phenix, please schedule a training session with Sam. Sam can also provide additional information on the associated cell culture facilities and operating procedures.
- Susan Hall (Susan_Hall@brown.edu) is the manager of the Opera Phenix imaging facility. Please contact Sue if you think the system needs to be serviced, if you'd like to have your name added to the mailing list, or if you need access to the online signup calendar.
- Kia Huffman (Kia_Huffman@brown.edu) is a Cyberinfrastructure Architect. Please contact Kia if you have questions about saving data or data management.
- Derek Merck (Derek_Merck@brown.edu, webpage) is the Director of Computer Vision and Image Analysis, DDI. Please contact Derek if you have questions about advanced image analysis or machine learning.
- 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.
Publication & Funding. For publications that made use of the Opera Phenix, please acknowledge Center funding: The Opera Phenix in the Center to Advance Predictive Biology 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. To get access to the calendar, please contact Susan Hall at Susan_Hall@brown.edu or 401-863-2985.
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.
When signing up on the calendar, please enter the following information:
- Title field: enter your first and last name (e.g. Eric Johnson)
- Description field: enter PI's last name (e.g. Smith)
Protocols, user manuals and application guides can be found on the password-protected 'protocol page'