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

Flow cytometry From basic theory to practical matters

Flow cytometry From basic theory to practical matters

Martial JAUME([email protected])

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


Overview of the optics systemOverview of the optics system

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

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/


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...

Documents

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