flow cytometry from basic theory to practical matters · flow cytometry from basic theory to...
TRANSCRIPT
![Page 1: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/1.jpg)
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
![Page 2: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/2.jpg)
A meaningful nameA meaningful name
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 3: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/3.jpg)
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
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 4: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/4.jpg)
Flow cytometerFlow cytometer
Background informationabout the instrument
![Page 5: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/5.jpg)
Flow cytometer: 3 subsystemsFlow cytometer: 3 subsystems
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 6: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/6.jpg)
System part 1: FluidicsSystem part 1: Fluidics
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 7: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/7.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
FluidicsFluidics
![Page 8: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/8.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Influence of the sheath flow rateInfluence of the sheath flow rate
![Page 9: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/9.jpg)
System part 2: Optics systemSystem part 2: Optics system
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 10: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/10.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Overview of the optics systemOverview of the optics system
![Page 11: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/11.jpg)
Overview of the optics systemOverview of the optics system
http://www.bd.com/videos/bdb/training/itf/optics/player.html
![Page 12: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/12.jpg)
Light collection through filtersLight collection through filters
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 13: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/13.jpg)
System part 3: ElectronicsSystem part 3: Electronics
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 14: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/14.jpg)
ElectronicsElectronics
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 15: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/15.jpg)
ElectronicsElectronics
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 16: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/16.jpg)
ElectronicsElectronics
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 17: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/17.jpg)
ElectronicsElectronics
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 18: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/18.jpg)
ElectronicsElectronics
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 19: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/19.jpg)
Starting experiment and analyzing dataStarting experiment and analyzing data
Practical considerations and examples of data acquired by flow cytometry
![Page 20: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/20.jpg)
Optical measurement: Light scatteringOptical measurement: Light scattering
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 21: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/21.jpg)
Optical measurement: Light scatteringOptical measurement: Light scattering
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 22: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/22.jpg)
Optical measurement: Light scatteringOptical measurement: Light scattering
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 23: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/23.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Optical measurement: FluorescenceOptical measurement: Fluorescence
![Page 24: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/24.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Optical measurement: FluorescenceOptical measurement: Fluorescence
![Page 25: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/25.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Optical measurement: FluorescenceOptical measurement: Fluorescence
![Page 26: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/26.jpg)
Excitation and Emission SpectraExcitation and Emission Spectra
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
For further details visit http://www.bdbiosciences.com/spectra/
![Page 27: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/27.jpg)
Multiple colors and spilloverMultiple colors and spillover
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 28: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/28.jpg)
Spillover and signal compensationSpillover and signal compensation
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 29: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/29.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Spillover and signal compensationSpillover and signal compensation
![Page 30: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/30.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Spillover and signal compensationSpillover and signal compensation
![Page 31: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/31.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Spillover and signal compensationSpillover and signal compensation
![Page 32: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/32.jpg)
Data collection and displayData collection and display
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 33: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/33.jpg)
Population gatingPopulation gating
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 34: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/34.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Population gatingPopulation gating
![Page 35: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/35.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Population gatingPopulation gating
![Page 36: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/36.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Color gatingColor gating
![Page 37: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/37.jpg)
Chart flow of data collection/analysisChart flow of data collection/analysis
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 38: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/38.jpg)
Analyzing histogram plotAnalyzing histogram plot
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 39: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/39.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Analyzing histogram plotAnalyzing histogram plot
![Page 40: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/40.jpg)
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
Analyzing dot plotAnalyzing dot plot
![Page 41: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/41.jpg)
Applications of flow cytometryApplications of flow cytometry
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 42: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/42.jpg)
ImmunityImmunity
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 43: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/43.jpg)
CancerCancer
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 44: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/44.jpg)
Molecular biologyMolecular biology
http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.html
![Page 45: Flow cytometry From basic theory to practical matters · Flow cytometry From basic theory to practical matters Flow cytometry From basic theory to practical matters Martial JAUME](https://reader034.vdocuments.site/reader034/viewer/2022043018/5f3a902b51fd2809475aa186/html5/thumbnails/45.jpg)
Want more…??Want more…??
Interpreting flow cytometry data: a guide for the perplexed.Herzenberg, L. A. et al.Nat Immunol 7, 681-685. (2006)..