Platform

Flow Cytometry

The flow cytometry core facility allows multiparametric analysis of cell populations as well as particles with specific physical characteristics.

Some devices also make it possible to perform sorting where cells are physically separated in subpopulations according to specific criteria. The sorted cells (purified) can then be used in subsequent experiments.

Contact
Annie Gosselin
Phone (office)
(514) 343-6111, ext. 8094
Email
annie.gosselin.2@umontreal.ca

Services offered:

  • Sterile cell sorting: 1 to 4 ways, in tubes, plates or on slides
  • BL2+ compatible and Human cell compatible sorting
  • Training available for acquisition and analysis by the flow cytometry core facility staff
  • Supervised acquisition/analysis
  • Self-service acquisition/analysis (for experienced or frequent users)
  • High-throughput acquisition/screening (96- and 384-well plates)
  • Access to analysis station with DIVA, FlowJo, ModFit LT, ForeCyt, Everest softwares
  • Advice on choice of fluorochromes and protocols for many applications
  • Consulting in development of new applications of flow technology

Note: The operation of our cell sorting system is restricted to Flow Cytometry Core Facility personnel.

Equipment

BD FACS Celesta cell analyser

The Celesta comes with 3 lasers which allow the detection of up to 14 parameters and 12 fluorochromes, and a high-throughput module (HTS) for cell screening into 96- and 384-well plates.

  • Blue laser 488 nm: 5 PMT detectors (SSC + 4 fluorochromes*) and a FSC photodiode detector
    *Possibility of multiple combinations of 4 filters among the following :
    750LP 780/60 (ex.: PE-Cy7)
    670LP 695/40 (ex.: PerCP, PerCP-Cy5.5, 7-AAD)
    635LP 670/30 (ex.: PE-Cy5)
    600LP 610/20 (ex.: PE-CF594, PE-TexasRed, PI)
    550LP 575/25 (ex.: PE)
    505LP 530/30 (ex.: FITC, Alexa488, GFP)
  • Red laser 640 nm: 3 PMT detectors
    750LP 780/60 (ex. : APC-Cy7)
    690LP 730/45 (ex.: Alexa700)
    670/30 (ex. : APC, Alexa647)
  • Violet laser 405 nm: 5 PMT detectors
    750LP 780/60 (ex.: BV786)
    655LP 670/30 (ex.: BV650)
    595LP 610/20 (ex.: BV605)
    505LP 525/50 (ex.: BV510, V500, AmCyan, Ametrine)
    450/40 (ex. : BV421, PacificBlue, V450)

More information about BD FACS Celesta cell analyser.

 

BD FACSCantoII cell analyser

The CantoII comes with 3 lasers which allow the detection of up to 10 parameters and 8 fluorochromes. Automated acquisition of tubes is made possible by the carousel loader option (up to 40 tubes per carousel).

  • Blue laser 488 nm: 5 PMT detectors (SSC + 4 fluorochromes*) and a FSC photodiode detector
    *Possibility of multiple combinations of 4 filters** among the following:
    735LP 780/60 (ex.: PE-Cy7)
    655LP 670LP (ex.: PE-Cy5, PE-Cy5.5, PerCP, PerCP-Cy5.5, 7-AAD)
    556LP 585/42 (ex.: PE, PI)
    502LP 530/30 (ex.: FITC, Alexa488, GFP)
    Simultaneous reading of YFP and GFP:
    530LP 550/30 (YFP)
    500LP 510/25 (GFP)
    mCherry detection (only for samples with high intensity signal):
    556LP 605/40 (mCherry)
  • Red laser 633 nm : 2 PMT detectors*
    *Possibility of multiple combinations of 2 filters** among the following :
    735LP 780/60 (ex.: APC-Cy7)
    660/20 (ex.: APC, Alexa647)
    690LP 710/40 (ex.: Alexa700)
  • Violet laser 405 nm: 2 PMT detectors*
    *Possibility of multiple combinations of 2 filters** among the following :
    502LP 510/50 (ex.: BV510, V500, AmCyan, Ametrine)
    450/50 (ex.: BV421, PacificBlue, V450)
    485/22 (ex.: CFP)
    735LP 780/60 (ex.: BV786)
    690LP 710/40 (ex.: BV711)
    595LP 660/20 (ex.: BV650)
    595LP 610/20 (ex.: BV605)

More information about BD FACSCantoII cell analyser.

Bio-Rad ZE5 (Yeti) cell analyser

The ZE5 comes with 5 lasers which allow the detection of up to 29 parameters and 26 fluorochromes. It has a high-throughput (HTS) reading option and can accommodate samples in eppendorf tubes, 5mL tubes and in 96- and 384-well plates. A small particle detector (SPD) allows the analysis of microparticules <0.3um. The Everest software can be installed on any personal computer and allows preparing the experimental setup prior to going to the FACS as well as doing post-acquisition analysis.

  • Blue laser 488 nm: 7 PMT detectors (SSC + 6 fluorochromes), and a PMT FSC 488/10
    488/10 (SSC)
    509/24 (ex.: FITC)
    549/15 (ex.: YFP)
    583/30 (ex.: DyeCycle Orange)
    615/24 (ex.: Propidium iodide)
    670/30 (ex.: PerCP)
    710/20 (ex.: PerCP-eFluor710)
  • Yellow-green laser 561 nm : 7 PMT detectors
    577/15 (ex.: PE)
    589/15 (ex.: mStrawberry)
    615/24 (ex.: Pe-TexasRed)
    640/20 (ex.: AlexaFluor610)
    670/30 (ex.: PE-Cy5)
    720/60 (ex.: PE-Cy5.5)
    750LP (ex.: PE-Cy7)
  • Red laser 640 nm : 3 PMT detectors
    670/30 (ex.: APC)
    720/60 (ex.: Alexa700)
    750LP (ex.: APC-Cy7)
  • Violet laser 405 nm : 7 PMT detectors, and a PMT FSC 405/10 (SPD)
    420/10 (ex.: BV421)
    460/22 (ex.: V450)
    525/50 (ex.: BV510)
    615/24 (ex.: BV605)
    670/30 (ex.: BV650)
    720/60 (ex.: BV711)
    750LP (ex.: BV786)
  • UV laser 355 nm: 3 PMT detectors
    447/60 (ex.: DAPI) or 387/11 (ex.: BUV395)
    525/50 (ex.: BUV496)
    650LP (ex.: BUV737)

More information about Bio-Rad ZE5 (Yeti) cell analyser.

IntelliCyt iQue cell screener

This device has 2 lasers and allows the detection of up to 6 parameters and 4 fluorochromes.
The analysis is done directly from samples into 96- or 384-well plates.

iQue cell screener characteristics:

  • No dead volume
  • Minimum acquisition volume: 1ul
  • Minimum well volume: 10ul
  • Minimum analysis time for a 96-well plate: 3 minutes
  • Minimum analysis time for a 384-well plate: 12 minutes
  • ForeCyt software can generate a heatmap of the plates for a rapid identification of the wells of interest.
  • Blue laser 488nm: 4 detectors (SSC + 3 fluorochromes)
    FL1 533/30 (ex.: FITC, Alexa488, GFP)
    FL2 585/40 (ex.: PE)
    FL3 670LP (ex.: PE-Cy5, PerCP-Cy5.5, PE-Cy7, 7-AAD)
  • Red laser 633nm: 1 detector
    FL4 675/25 (ex.: APC, Alexa647)

More information about IntelliCyt iQue cell screener

autoMACS cell separator

The autoMACS cell separator allows the separation by positive or negative selection of a sub-population of cells of interest using magnetic immuno-labelling kits available from Miltenyi company. Various separation programs are optimized for different applications.

  • Technical characteristics:
    Programs: 9 standard programs
    Lenght of the programs from 2.5 to 10 minutes per ml of sample
    Speed: 10e7 cells / second per ml of sample
    Column capacity: 2 x 10e8 stained cells and 4 x 10e9 total nucleated cells.

More information about autoMACS cell separator

BD FACSAria cell sorters (2)

The two cell sorters have the following capabilities:

  • From 1- to 4-way cell sorting
  • Sorting in plates (6, 12, 24, 48, 96 or 384 wells)
  • Sorting on slides
  • The samples to be sorted can be provided in 1ml, 5ml or 15ml conical bottom tubes.
  • Choice of collection tubes: 0.5ml, 1ml, 1.5ml Eppendorf tubes, 5ml or 15ml conical bottom tubes.
  • Adjustable temperature of the sample chamber (4ºC, 20ºC, 37ºC or 42ºC
  • Adjustable temperature for the collection chamber for 5ml or 15ml tubes (2ºC to 50ºC) 70µm, 85 µm and 100µm nozzle sizes
  • Cell sorting at a range of pressures adapted to cell size and fragility
  • Cell sorter under a biological hood for potentially infectious sorting and ensure sterile sorting for downstream cell culture.
  • Additional filters to detect fluorochromes other than those listed are available.

More information about BD FACSAria cell sorters

BD FACSAria IIIu cell sorter

This cell sorter has 3 lasers, giving the possibility to sort cells on the basis of the analysis of up to 14 parameters (12 fluorochromes).

  • Blue laser 488 nm: 5 PMT detectors (SSC + 4 fluorochromes) and a FSC photodiode detector
    690LP 695/40 (ex.: PerCP-Cy5.5)
    655LP 675/14 (ex.: PerCP)
    600LP 610/20 (ex.: Red613, YFP)
    502LP 530/30 (ex.: FITC, Alexa488, GFP)
  • Yellow-Green laser 561 nm : 5 PMT detectors
    735LP 780/60 (ex.: PE-Cy7)
    685LP 710/50 (ex.: PE-Cy5.5)
    630LP 670/14 (ex.: PE-Cy5, 7-AAD, mPlum)
    600LP 610/20 (ex.: mCherry, RFP, PE-TexasRed, PI)
    582/15 (ex.: PE, Tomato)
  • Red laser 633 nm: 3 PMT detectors
    735LP 780/60 (ex.: APC-Cy7)
    685LP 710/40 (ex.: Alexa700)
    660/20 (ex.: APC, Alexa647)

BD FACSAria IIu cell sorter

This cell sorter has 3 lasers which allows the sorting of cells on the basis of the analysis of up to 13 parameters (11 fluorochromes).

  • Blue laser 488 nm: 7 PMT detectors (SSC + 6 fluorochromes) and a FSC photodiode detector
    735LP 780/60 (ex.: PE-Cy7)
    685LP 695/40 (ex.: PerCP-Cy5.5, PE-Cy5.5)
    655LP 670/14 (ex.: PE-Cy5, PerCP, 7-AAD)
    600LP 610/20 (ex.: PE-TexasRed, PI)
    556LP 575/26 (ex.: PE)
    502LP 530/30 (ex.: FITC, Alexa488, GFP)
    Filters can be changed to simultaneously read YFP and GFP :
    530LP 550/30 (YFP)
    500LP 510/25 (GFP)
  • Red laser 633 nm: 3 PMT detectors
    735LP 780/60 (ex.: APC-Cy7)
    690LP 710/40 (ex.: Alexa700)
    660/20 (ex.: APC, Alexa647)
  • Violet laser 405 nm : 2 PMT detectors*
    *Possibility of multiple combinations of 2 filters** among the following :
    735LP 780/60 (ex.: BV786)
    690LP 710/40 (ex.: BV711)
    595LP 660/20 (ex.: BV650)
    595LP 610/20 (ex.: BV605)
    502LP 530/30 (ex.: BV510, V500, AmCyan , Ametrine)
    450/40 (ex.: BV421, PacificBlue, V450)
    485/22 (ex.: CFP)

Staff