Flow Cytometry Instruments
The facility has several Becton Dickinson (BD) analyzers. Please note that these analyzers differ in the types of lasers they are equipped with and also the powers of the lasers.
Each instrument has its own default configuration and was designed for a specific set of commonly used fluorochromes in which all detectors are used simultaneously. Filters in this configuration may not necessarily be optimal for your particular fluorochrome combination. Additional filters are available to users and can be signed out from the FCF office. The instruments must be returned to their default configuration after use. See FCF staff for assistance on choosing optimal filters. Please note that the fluorochrome list is not exhaustive. Other fluorochromes not listed may also be detected.
The LSR II SC and LSR II GC have a High Throughput Sampler (HTS) option. The HTS unit, which mounts onto the instrument, allows for acquisition from 96- or 384-well plates. Additional training is required to use the HTS option.
For every appointment, clients must indicate the Risk Group categories of any biological agents they contain. The FCF operates in Containment Level 2 (CL2) Facilities and can handle samples designated as RG1 or RG2. No samples or agents designated as RG3 and RG4 can be brought into FCF labs. More information on our biosafety policies is available in our FCF-iLab.
Click on the instruments below for more information about each instrument including instrument configurations.
For information on access please see training. More information on technical specifications for these instruments can be obtained from the FCF manager.
|Instrument||LSR II CFI||LSR Fortessa||LSR II GC||LSR II OICR||LSR II SC|
|Location||PGCRL, 18.9430||PGCRL, 18.9430||PGCRL 18-9430||PMCRT, 2-504||PMCRT, 2-504|
|# of colours||15||17||12||18||14|
|Violet||25 mW||50 mW||25 mW||50 mW||50 mW|
|Blue||50 mW||100 mW||100 mW||50 mW||20 mW|
|Green||100 mW (552 nm)||150 mW (532 nm)||-||200 mW (532 nm)||-|
|Red||40 mW||40 mW||20 mW||40 mW||40 mW|
Additional filters are available for BD instruments and can be signed out from the FCF office. Please submit a “General Assistance” request (listed under Special Assistance category) in iLab to request alternate filter (s). A member of our staff will assist you to sign out the filter(s) you require during normal working hours (Monday to Friday, 10 a.m. to 6 p.m.). Please return the filter(s) to a member of the our staff at the end of your appointment.
If your appointment occurs after hours or on the weekends, the filter(s) must be returned by 10 a.m. the next working day. Please note that instruments must be returned to their default filter configuration at the end of your appointment. See FCF staff for assistance on choosing optimal filters.
|Filters for detecting GFP and YFP simultaneously||Quantity|
|550/30 BP (for YFP)|
510/21 or 510/50 BP (for GFP) 525 LP
The facility houses four FACSArias (BD), two MoFlos (Beckman Coulter), one SH800 (Sony) and one Influx (BD). All of our sorters are located in biosafety cabinets for sorting human samples and other biohazardous samples.
For every sort appointment clients must indicate the types of samples to be sorted and identify the Risk Group categories of any biological agents they contain. The FCF operates in Containment Level 2 (CL2) Facilities and can handle samples designated as RG1 or RG2. No samples or agents designated as RG3 and RG4 can be brought into FCF labs. Most sorters are operated exclusively by our staff, with the exception of the Sony SH800.
Click on the instruments below for more information on configurations.
BD Cell Sorters
|Instrument||Aria II RITT||Aria III CFI||Aria Fusion A||Aria Fusion B||Influx|
|Location||PMCRT, 2-504||PMCRT, 2-203||PMCRT, 2-504||PMCRT, 2-203||PMCRT, 2-504|
|# of colours||11||18||16||16||9|
|Restricted||FCF staff operated||FCF staff operated||FCF staff operated||FCF staff operated||FCF staff operated|
|Violet||15 mW||10 mW||85 mW||85 mW||50 mW|
|Blue||20 mW||20 mW||50 mW||50 mW||200 mW|
|Green||-||50 mW (561 nm)||150 mW (532 nm)||150 mW (532 nm)||-|
|Red||18 mW||15 mW||100 mW||100 mW||30 mW|
Beckman Coulter Cell Sorter
|# of colours||11-14|
|Restricted||FCF staff operated|
* Note: The violet and UV laser cannot be used simultaneously on the MoFlo XDP.
Sony Cell Sorters
Magnetic Cell Separators
Magnetic cell separation technology can be used to enrich heterogeneous cell samples for particular cell populations prior to sorting, or as an endpoint itself (e.g. positive selection of CD11c+ DC, generation of Lineage negative cells for adoptive transfer).
Our facility has an automated magnetic separator, autoMACS Pro Separator (Miltenyi Biotec), located in room 2-504 at PMCRT. A mandatory 15-minute training session provided by FCF staff is a prerequisite.
autoMACS Pro Separator
Location: PMCRT 2-504
The autoMACS Pro Separator is a bench-top automated magnetic cell separator used for cell isolation and enrichment. Up to six samples can be processed automatically in volumes ranging from 0.2 mL to 50 mL. Preset cell separation programs allow positive or negative selection of magnetically labeled cells using MACS columns.
Cell separation programs are chosen based on levels of antigen expression and desired levels of recovery and purity. Cells are first labeled by the user with antibodies and/or MACS magnetic microbeads and then loaded onto the autoMACS where cells are passed through one or two columns (depending on the program). Both labeled and non-labeled fractions are eluted into tubes in chilled tube racks. Wash programs are performed between separations. The unit is in a bio-safety cabinet for aseptic handling of samples.
The FCF operates in Containment Level 2 (CL2) Facilities and can handle samples designated as RG1 or RG2. No samples or agents designated as RG3 and RG4 can be brought into FCF labs. More information on our biosafety policies is available in our FCF-iLab
- 10 standard programs
- Program duration: 2.5-10 minutes per sample mL, 4 minutes per wash
- Maximum number of cells: 2 x 108 labeled cells, 4 x 109 total cells per separation
- Minimum number of cells: 1 x 105
- Sample volume: 200 µl to 50 mL
- Running buffer: can be purchased from Miltenyi (PBS, 2 mM EDTA, 0.5% BSA, 0.09% NaAzide, cat#130-091-221, $330 for 6 x 1.5L) or make your own in 1L pyrex bottles
- MACS microbeads and antibodies for labeling