Flow Cytometry Core
Facility (FCCF)

The facility is equipped with five analyzers and two cell sorters.

ANALYZERS:

BD FACSCanto II

• 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

CYTEK Aurora

• 4 lasers full spectrum analyzer (405/488/561/633 nm); 48parameters
   

SORTERS:

BD FACSAria Fusion

4 lasers (405/488/561/633 nm); 18 parameters cell sorter with biosafety cabinet*
* The cell sorter is operator based. You can book an appointment on Infinity X (see booking calendar tab). The appointment request will be confirmed or rescheduled based on staff availability.

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.

SONY SH800

• 4 lasers (405/488/561/633 nm); 8 parameters; 70 uM or 100uM sorting chips offered as options for the sorts.

Beckman Coulter CytoFlex SRT

• 4 lasers (405/488/561/638 nm); 17 parameters; **instrument located in the GCI BSL-3 facility in a biosafety cabinet**

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.

Sample Collection

Cells can be collected in various types of tubes or plates:

• Eppendorf tubes
• 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.
 

To gain access to the facility and receive their initial training, new users will need to create their account on McGill’s scientific platforms administration system, Infinity X of which the FCCF is part of. Users can access the software using the link below:

Infinity X

A recorded video explaining login procedure and how to navigate the software can be found here: Infinity explaining video.

Monthly billing by the FCCF will be based on calendar booking time. Users need to book the right amount of time needed on the instrument and adjust the actual used time afterwards on Infinity.

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.

Tel Office: 514-398-8203
Tel Lab: 514-398-4400 ext.00809

Address
McGill University
Life Science Complex
Bellini building,
Room 337a (Office)
Room 343 (Sorters)
Room 345 (Analyzers)
3649 Promenade Sir William Osler
Montréal, Qc, Canada, H3G 0B1

Hours of operations:
Monday - Friday: 9:30 AM - 5:30 PM
After Hours: The cytometers are accessible after hours for trained users.