Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to...

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Flow cytometry From basic theory to practical matters

Martial JAUME(breizh@hku.hk)

HKU-Pasteur Research Centre

Flow cytometry workshopHKU, 21-22 October 2010

A meaningful nameA meaningful name

http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html

DefinitionsDefinitions

Flow cytometry is a powerful technology for characterizing and analyzing cells. It simultaneously measures and analyzes multiple physical characteristics of single particles, usually cells, as they move in a fluid stream through a beam of light

Flow cytometerFlow cytometer

Background informationabout the instrument

Flow cytometer: 3 subsystemsFlow cytometer: 3 subsystems

System part 1: FluidicsSystem part 1: Fluidics

FluidicsFluidics

Influence of the sheath flow rateInfluence of the sheath flow rate

System part 2: Optics systemSystem part 2: Optics system

Overview of the optics systemOverview of the optics system

http://www.bd.com/videos/bdb/training/itf/optics/player.html

Light collection through filtersLight collection through filters

System part 3: ElectronicsSystem part 3: Electronics

ElectronicsElectronics

Starting experiment and analyzing dataStarting experiment and analyzing data

Practical considerations and examples of data acquired by flow cytometry

Optical measurement: Light scatteringOptical measurement: Light scattering

Optical measurement: FluorescenceOptical measurement: Fluorescence

Excitation and Emission SpectraExcitation and Emission Spectra

For further details visit http://www.bdbiosciences.com/spectra/

Multiple colors and spilloverMultiple colors and spillover

Spillover and signal compensationSpillover and signal compensation

Data collection and displayData collection and display

Population gatingPopulation gating

Color gatingColor gating

Chart flow of data collection/analysisChart flow of data collection/analysis

Analyzing histogram plotAnalyzing histogram plot

Analyzing dot plotAnalyzing dot plot

Applications of flow cytometryApplications of flow cytometry

ImmunityImmunity

CancerCancer

Molecular biologyMolecular biology

Want more…??Want more…??

Interpreting flow cytometry data: a guide for the perplexed.Herzenberg, L. A. et al.Nat Immunol 7, 681-685. (2006)..

Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to...

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