Flow Cytometry

 Brochure PDF

Flow cytometry is a method for measuring and analysing the physical characteristics of individual cells or particles. It consists of running single cells in suspension in front of one or more laser beams and then collecting fluorescent signals emitted by each cell as well as the diffused light. Data collected are translated into graphs or histograms and statistics concerning cell populations and parameters under study (%, CV, Fluorescence Intensity, etc.). As well, sorting instruments may be used to physically isolate cell subsets according to specific criteria and these may be used in other types of experiments.

The principal applications of flow cytometry are:

• immunophenotyping
• cell cycle, apoptosis study
• cell proliferation analysis
• intracellular cytokines analysis
• signal transduction pathways study (phospho-proteins)
• gene expression measurement with fluorescent proteins (GFP, YFP, etc.)
• intracellular Ca²⁺, intracellular pH measurement

IRIC’s Flow Cytometry Core Facility offers high quality, cost-effective services primarily to IRIC/Université de Montréal investigators in the course of their research initiatives. It is also accessible to outside scientists from both academia and industry through collaborative and contract research agreements. Our staff is available for project consultations and can train users on equipment operation.

The facility is located on the first floor of IRIC’s Marcelle-Coutu Pavilion, alongside our Bio-Imaging Centre.

Overview of Services

IRIC’S Flow Cytometry Facility offers the following services:

• Cell sorting: 1 to 4 ways, in tubes, plates or on slides
• Supervised acquisition/analysis
• Unsupervised acquisition/analysis (for experienced or frequent users)
• Analysis station with DiVa, FlowJo, FCS Express3, ModFit LT
• Training in acquisition and analysis
• Advice on choice of fluorochromes and protocols for many applications
• Consulting in development of new applications of flow technology

The operation of our cell sorting equipment is restricted to properly trained facility personnel.

Instrumentation

Our state-of-the-art sorting and acquisition/analysis equipment includes a BD LSR II flow cytometer and BD FACSAria high-speed cell sorters that feature completely redesigned optics with improved signal detection sensitivity and channel resolution. These instruments are equipped with an advanced digital electronic system allowing for faster signal processing. With DiVa’s digital software, automatic compensation for all fluorescent signals is available, making it easy to conduct multicolour experiments. The autoMACS is an automated bench-top magnetic sorting system capable of sorting millions of cells per second for positive selection as well as depletion.

BD LSR II Flow Cytometer
The BD LSR II can be configured with up to 3 lasers and the ability to detect up to 12 parameters and 10 colours:

• 488 nm: 7 detectors (SSC + 6 colours)
• 633 nm: 2 detectors
• 355 nm (UV): 2 detectors

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BD FACSAria Cell Sorters
IRIC’S two BD FACSAria high-speed cell sorters provide the following options:

• 1 to 4 ways sorting
• Sorting in plates
• Sorting on slides
• Sample sorting in 1ml tube, 5ml tube or 15 ml tube with conical bottom
• Collection tubes: 1ml, 5ml or 15 ml with conical bottom
• Controlled temperature for the bulk injection chamber (samples) (4ºC, 20ºC, 37ºC or 42ºC)
• Controlled temperature for the collection tubes of 5ml or 15ml (2ºC to 50ºC)
• 70µm and 100µm nozzles
• Sorting at various pressures according to cell size and fragility

BD FACSAria Cell Sorter # 1
With its 3 lasers, this FACSAria can detect up to 13 colours, for a total of 15 parameters:

• 488 nm: 8 detectors (SSC + 7 fluorochromes)
• 633 nm: 3 detectors
• 407 nm (violet): 3 detectors

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BD FACSAria Cell Sorter #2
With its 2 lasers, this instrument can detect up to 10 colours, for a total of 12 parameters:

• 488 nm : 8 detectors (SSC + 7 colors)
• 633 nm : 3 detectors

AutoMACS Separator
The autoMACS separator—an automated cell separator and sorting system—features several programmes to optimize the sorting of each cell preparation. For example, by using the positive selection programmes, the autoMACS separator is capable of isolating up to 2 X 108 pure target cells within minutes.
Specifications:

• Programs: 9 standards
• Program duration: 2.5-10 minutes per sample ml
• Speed: 10⁷ cells/second per sample ml
• Column capacity: 2 x 10⁸ magnetically labelled cells, 4 x 10⁹ total nucleated cells

Pricing

 Consult our current rates.

Contacts

For more information on our Flow Cymometry Core Facility or to schedule an appointment, please contact:

Danièle Gagné
T + 514 343.6111 ext. 1.8094
daniele.gagne@umontreal.ca

Claude Perreault, M.D., F.R.C.P.(C)

T + 514 343.6126
claude.perreault@umontreal.ca

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