Welcome to the Flow Cytometry Core Facility (FCCF)!
Located in the Bellini Pavilion of the Life Science Complex, the Core facility provides instrumentation and technical assistance to perform laser-based flow cytometric analysis, as well as providing an operator based cell sorting service.
Please follow this link to access our external webpage for a complete description of our services, instrument details, available resources and core policies:
- Services and Fees
- Cell Sorting and Sample Preparation
- Booking Calendar
- Contact Us
- Acknowledging Core Support
Services and Fees
The Flow Cytometry Core Facility is equipped with 5 different flow cytometers for various applications. An operator-based cell sorting service is also offered. You can refer to the equipment section of this website to view the instrument available. For a more complete description of the equipment (configurations and technical specificity), please consult our external webpage (https://mimmflow.weebly.com/).
We provide the mandatory training for the users to gain access to the cytometers. We offer consultation for experiment design, data analysis and other technical assistance. Users or clients that won’t be doing regular acquisition and analysis can also ask for operator assisted data acquisition and analysis. We also provide dongles for the analysis software FlowJo. They can be rent per day by any users.
The facility is open Monday through Friday from 9:30 a.m. until 5:00 p.m. for cell sorting services and user assistance. The cytometers are accessible at any time for trained users.
Contact us for an instrument training, to schedule a sort or for technical advice on experimental design.
|BD FACSCanto II||$30/hour||$40/hour||$45/hour||$100/hour|
|BD LSR Fortessa||$30/hour||$40/hour||$45/hour||$100/hour|
|Additional Assisted Fees||$50/hour||$50/hour||$50/hour||$50/hour|
*Life Science Complex (LSC) rate: McIntyre building labs, GCRC labs, Bellini building labs, Stewart building labs.
Monthly bill is capped at $650 for analyzers (LSC members only).
The facility is equipped with five analyzers and two cell sorters.
• 2 lasers (488/633 nm); 6 parameters
BD FACSCanto II (2X)
• 3 lasers (405/488/633 nm); 10 parameters
• 2 lasers (488/633 nm); 8 parameters
BD LSR Fortessa (2X)
• 5 lasers (355/405/488/561/640 nm); 20 parameters
• 4 lasers (405/488/561/633 nm); 18 parameters
BD FACSAria Fusion
4 lasers (405/488/561/633 nm); 18 parameters cell sorter with biosafety cabinet*
* The sorter is operator based. You can contact the staff to schedule a sort. Different time slots will be offered to you based on instrument and staff availability. Note that given current demand for cell sorting appointments, we recommend that you book your sort at least 2 weeks ahead of time.
BD FACS Aria III
3 lasers (405/488/640 nm); 13 parameters cell sorter equipped with a biobubble*
*The policies are the same for this sorter. The biobubble is a cabinet that exceeds Class 1 Biological safety specifications.
Cell Sorting and Sample Preparation
Cell Sorting and Sample Preparation
The sort can be performed at different pressure and nozzle size. We offer 4 different sorting conditions for various cell types:
-70 uM /70 PSI (Splenocytes/Thymocytes/PBMC/Whole blood/bone marrow)
-85 uM /45 PSI (Transduced-transfected splenocytes, thymocytes, bone marrow cells / B or T cell lines)
-100 uM /20 PSI (Fibroblasts, cultured primary cells, medium to large cell lines, endothelial cells, adherent cell lines, transfected cell lines)
-130 uM /16 PSI (Dissociated tumor cells/ large cell lines/ delicate- sensitive cells)
Please consult facility staff to decide the ideal conditions for your sort.
Note that the nozzle size and sample concentration will affect the time of your sort. The optimal threshold rate to achieve the best recovery/efficiency ratio will be the determining factor.
Basic Sorting buffer
Prepare the following buffer in which to suspend your samples after staining, prior to cell sorting:
1X Phosphate Buffured Saline (PBS) or Hanks Balanced Salt Solution (HBSS) (free of Ca2+ or Mg2+)
1 % heat-inactivated FBS (or BSA)
1 mM EDTA (can be removed for simple lymphocyte populations)
Sterilize with a 0.2uM filter and store the solution at 4°C.
Before bringing your samples to be processed, filter them through a 40/70 uM mesh filter. Consider bringing extra buffer for dilution purposes.
Modifications for specific cell types:
For Sticky Cells: The EDTA concentration can be increased to 5 mM. Note that cell tolerance for EDTA varies; too much can kill your cells. FBS can neutralize adherent cells. Consider using cell dissociation products that maintain cell suspensions.
For sensitive cells: You can add 25 mM HEPES to stabilize the pH.
For samples with high cell death: DNA will be released in solution by the dead/dying cells. This significantly increases cell clumping. Add DNAse to your sample prior to sorting.
Cells can be collected in various types of tubes or plates:
-5 ml FACS tubes (ideally polypropylene as cells adhere less than in polystyrene)
-15 ml conical tubes (2-way sort only)
-Multi-well plates (6, 12, 24, 48, 96, 384) (1-way sort only)
-Slides (1-way sort only)
Coat the recovering tube/plate with media or PBS before arriving. You can add roughly 10-20% of the receptacle volume (e.g. 2ml in a 15ml conical tube). Consider adding a higher concentration of FBS to the recovery media for sensitive samples.
The collection tubes can also be kept at 4°C during the sort. You can decide the collection temperature on the day of your sort and tell the operator.
After receiving proper training on a cytometer, the users will be granted card access to the facility rooms and will receive a login and password for the online calendar system. They will then be able to book a time-slot for an acquisition on an instrument they have been trained on.
Out of respect to the other users and to maintain reasonable instrument access for every user, make sure you book only the time needed for your experiment to avoid unused blocked-time.
Here is the link to the online calendar: http://lscflow.bookmylab.com
Directors of the Core
Dr. Jörg Fritz
Dr. Judith Mandl
Dr. Corinne Maurice
Dr. Martin Richer
Dr. Michel Tremblay
julien.leconte [at] mcgill.ca
camille.stegen [at] mcgill.ca
Tel Office: 514-398-8203
Tel Lab: 514-398-4400 ext.00809
Life Science Complex
Bellini building, room 337
3649 Promenade Sir William Osler
Montréal, Qc, Canada, H3G 0B1
Hours of operation
Monday - Friday: 9:30 AM - 5:30 PM
After Hours: The cytometers are accessible after hours for trained users.
Acknowledging core support
All users and investigators can help us maintain our excellence by acknowledging core support. When preparing manuscripts that contain work originating from services or resources provided by the Flow Cytometry Core Facility, please acknowledge support.
Here is a suggested acknowledgment:
“The flow cytometry work/ cell sorting was performed in the Flow Cytometry Core Facility for flow cytometry and single cell analysis of the Life Science Complex and supported by funding from the Canadian Foundation for Innovation.”
Please inform us of each publication acknowledging our core to help us combine and summarize them for grant application purposes.