Centre for Flow Cytometry and Microscopy
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Training courses

1. FACS training

Course details

FACS training is divided into the following components:

  • introductionIntroduction to flow basics which covers components of a flow cytometer and basics of fluorescence absorption and emission
  • useUse of FACSCalibur instrumentation, data acquisition and analysis
  • handsHands-on practice session using beads or prepared sample.

We use FACSCalibur analyzers equipped with two lasers, Macintosh G4 and BD CellQuest Pro data acquisition and analysis software.

Training schedules depend on instrument availability. Courses generally occur twice a month and are filled on a first-come, first-serve basis. Each training session takes three to fourhours and is split up into one- or two-day sessions to allow for sample preparation. At the end of the course, users may need to take advantage of assisted FACS sessions to complete their learning.

Requirements

It is important that you understand your project objectives and the biology of your cells. You will not succeed in adopting flow as a technological tool if you don't understand why you need to look at cells with flow cytometry.

Recommendations

We encourage you to bring the prepared cells or samples that you will most likely be working on for your project. Examples include GFP expression, DNA content or cell-surface immunofluorescence.

It is not advisable to register for the course if you do not have a sample or a FACS project ready (this includes antibodies and reagents).

2. LSRIILSR II and FACSCanto training

Course details

LSRIILSR II and FACSCanto training is divided into the following components:

  • advancedAdvanced topics such as filter optimization, instrument configuration, bi-exponential display and digital data
  • FACSDiva software
  • instrumentInstrument setup and usage
  • qualityQuality control
  • acquisitionAcquisition and analysis.

Training is available to users who are already experienced with flow via the FACSCalibur. If you do not have FACSCalibur training but absolutely require a panel using more than 4 fluorophores for your flow cytometry characterization, then training will be provided. Both the LSR II and FACSCanto use FACSDiva software, which is very different from CellQuest Pro. Digital flow cytometry requires due diligence in preparing the cytometer via single stained compensation controls and assigning appropriate PMT voltages. We focus on drawing your attention to improved quality control practices.

Some applications for which you can use the LSR II system include:

  • multi-fluor assays (14 parameter)
  • calcium flux assays
  • phosphorylated protein assays
  • H0342 side population assays
  • Simultaneous CFP-GFP-YFP-dsREd measurements

Recommendations

An excellent understanding of your application or the biology of your cells is recommended. This will help you use the software effectively.