Flow Cytometry Core Facility
Phong Le, PhD, Core Facility Director
ple@luc.edu
Bert Ladd, Core Facilities Manager
rladd@luc.edu
CBCC Room 212
Corbin Pomykata, Flow Cytometry Specialist
cpomykata@luc.edu
The FACS Core Facility occupies a 500 ft laboratory on the second floor of the Cancer Center. The Facility contains a FACSAria, a FACS Canto II, an LSRFortessa and an Amnis ImageStreamX Mark II. The FACSAria IIIu is equipped with 5 lasers – 405, 488, 561, 633, and near UV and is capable of detecting 15 simultaneous fluorochromes; the FACSCanto II bench-top flow cytometer equipped with 3 lasers – 405, 488, and 633 and is 8 color capable; and an LSRFortessa flow cytometer equipped with 4 lasers – 405,488, 561, and 633 and is 15 color capable. The Amnis ImageStreamX Mark II is equipped with 3 lasers – 405, 488, and 633 and is 6 color capable. The Chromium Controller is a chip technology that uses microfluidics to perform single cell partitioning and barcoding in minutes. The Chromium Controller enables integrated analysis of single cells at large scale. The Cytek Aurora is a full spectrum technology which provides the use of a wide array of fluorochromers without instrument reconfiguration This high resolution cytometer deliver high resolution data at the single-cell level and can resolve challenging issues such as high autofluorescence and low level of expresson of biomarkers.
Services
Consultation
- Basic consultation on experiment/panel design is available free-of-charge.
- In-depth design work will incur the hourly Consultation/Analysis charge at the discretion of core staff.
Data Analysis
- PC Workstations with analysis software (FlowJo, FCAP Array, etc.) are available free-of-charge, but time must be reserved on the portal.
- Basic consultation on analysis is available free-of-charge.
- In-depth analysis or analysis consultation will incur the hourly Consultation/Analysis charge at the discretion of core staff.
Data Management
- Data acquired in the facility is uploaded to the Shared Drive (\\fshsd01\V03\ONChsd\Shared\FACS) the following business day. Email rladd@luc.edu to request access.
- Data acquired in the facility is backed up to a physical archive for redundancy.
- DISCLAIMER: The Shared Drive and physical archive are not intended to be used as primary data storage. Download your data and back it up to a separate location ASAP.
Staff-Operated Acquisition and Cell Sorting
- Users can request staff-operated acquisition of their samples. Must be approved by staff in advance of booking.
- Users can request cell sorting for isolation of cell populations. Must be approved by staff in advance, as this service is entirely staff-operated.
Training/Education
- Users can be trained to independently acquire data on all instruments except the cell sorters (AriaIIIu and Aurora CS), which is entirely staff-operated.
- LSR Fortessa (BD) 4-Laser Conventional Flow Cytometer
- Novocyte Quanteon (Agilent) 4-Laser Conventional Flow Cytometer
- Aurora (Cytek) 5-Laser Full-Spectrum Flow Cytometer
- Amnis ImageStream MkII (Cytek) 3-Laser Imaging Flow Cytometer
- Users can be trained on basic data analysis via FlowJo; consultation available for advanced analysis.
- An introductory lecture for new users of the facility is offered roughly every 6 weeks.
Pricing
Cell Sorting (staff operated only)
- $75 for 1st hour, then $38 per hour.
Consultation/Analysis
- Basic: free-of-charge
- In-Depth: $60 per hour
Data Acquisition
- Independent: $10 per 30 minutes, minimum charge $16
- Staff-Operated: $75 per hour
Training for independent data acquisition
- $150 flat fee
- Includes introductory lecture, hands-on training, and basic analysis training.
Instruments
Amnis ImageStream MkII (Cytek) 3-Laser Imaging Flow Cytometer
- 4 Lasers: 405nm, 488nm, 640nm, 785nm
- 6 channels for imaging: fluorescence and/or brightfield
- Imaging data output allows analysis of co-localization, internalization, etc.
Aurora (Cytek) 5-Laser Full-Spectrum Flow Cytometer
- 5 Lasers: 355nm, 405nm, 488nm, 561nm, 633nm
- 64 channels for fluorescence, up to ~40 fluorescent parameters simultaneously
- Automated loader for 96-well plates and 40-tube rack
Aurora CS (Cytek) 5-Laser Full-Spectrum Cell Sorter
- 5 Lasers: 355nm, 405nm, 488nm, 561nm, 633nm
- 64 channels for fluorescence, up to ~40 fluorescent parameters simultaneously
- Droplet-based isolation of up to 6 simultaneous cell populations based on scatter and fluorescence
Chromium X Controller (10X Genomics)
- Single-cell barcoding for downstream RNA sequencing
FACS AriaIIIu (BD) 4-Laser Conventional Cell Sorter
- 4 Lasers: 405nm, 488nm, 561nm, 633nm
- 15 channels for fluorescent parameters
- Droplet-based isolation of up to 4 cell populations based on scatter and fluorescence
LSR Fortessa (BD) 4-Laser Conventional Flow Cytometer
- 4 Lasers: 405nm, 488nm, 561nm, 633nm
- 15 channels for fluorescent parameters
Novocyte Quanteon (Agilent) 4-Laser Conventional Flow Cytometer
- 4 Lasers: 405nm, 488nm, 561nm, 633nm
- 20 channels for fluorescent parameters
- Photodetectors with high dynamic range – minimal voltage adjustment required
- Automated loader for plates (96-well etc.) and 40-tube rack
Training classes and one-on-one support are available for investigators, students, and staff to use the FACSCanto II and LSRFortessa for data acquisition. Dedicated computer workstations offering Flowjo, CellQuest, and ModFit LT software packages are available for data analysis.
The FACS Core Facility also houses a Miltenyi-Biotec AUTOMACS, a fully automated bench-top sorter for positive or negative magnetic cell separation. Users can perform sterile or non-sterile separations based on their experimental needs.
Phong Le, PhD, Core Facility Director
ple@luc.edu
Bert Ladd, Core Facilities Manager
rladd@luc.edu
CBCC Room 212
Corbin Pomykata, Flow Cytometry Specialist
cpomykata@luc.edu
The FACS Core Facility occupies a 500 ft laboratory on the second floor of the Cancer Center. The Facility contains a FACSAria, a FACS Canto II, an LSRFortessa and an Amnis ImageStreamX Mark II. The FACSAria IIIu is equipped with 5 lasers – 405, 488, 561, 633, and near UV and is capable of detecting 15 simultaneous fluorochromes; the FACSCanto II bench-top flow cytometer equipped with 3 lasers – 405, 488, and 633 and is 8 color capable; and an LSRFortessa flow cytometer equipped with 4 lasers – 405,488, 561, and 633 and is 15 color capable. The Amnis ImageStreamX Mark II is equipped with 3 lasers – 405, 488, and 633 and is 6 color capable. The Chromium Controller is a chip technology that uses microfluidics to perform single cell partitioning and barcoding in minutes. The Chromium Controller enables integrated analysis of single cells at large scale. The Cytek Aurora is a full spectrum technology which provides the use of a wide array of fluorochromers without instrument reconfiguration This high resolution cytometer deliver high resolution data at the single-cell level and can resolve challenging issues such as high autofluorescence and low level of expresson of biomarkers.
Training classes and one-on-one support are available for investigators, students, and staff to use the FACSCanto II and LSRFortessa for data acquisition. Dedicated computer workstations offering Flowjo, CellQuest, and ModFit LT software packages are available for data analysis.
The FACS Core Facility also houses a Miltenyi-Biotec AUTOMACS, a fully automated bench-top sorter for positive or negative magnetic cell separation. Users can perform sterile or non-sterile separations based on their experimental needs.