20100617 ps
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Flow cytometry allows for the high-speed analysis and sorting of single cells. It provides multiparameter measurements of individual cells as they pass through the flow cytometer, allowing characterization of cell size, granularity, fluorescence, and other properties. This characterization can provide information on cell composition, physiology, morphology, and more. Flow cytometry is useful for applications like selection of cells with desired traits, analysis of algal cultures, and screening of samples.
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Flow cytometry allows for the quantitative and qualitative analysis of cell populations by measuring physical and chemical characteristics of individual cells as they flow in a single file through a laser beam. Cells are labeled with fluorescent markers and passed through the flow cytometer which measures light scattering and fluorescence emission. Data is collected and analyzed using plots that display fluorescence parameters of individual cells. Flow cytometry has many clinical applications including immunophenotyping, stem cell analysis, and detection of rare cell populations in diseases like cancer.
An introduction to flow cytometry- Ashwini.R
The document provides an introduction to flow cytometry. It describes flow cytometry as a technique that allows simultaneous multiparametric analysis of physical and chemical characteristics of single cells suspended in a fluid stream. Key components of a flow cytometer include fluidics, optics, detectors, and electronics. Cells are hydrodynamically focused into a single file stream and pass through a laser beam, where light scattering and fluorescence emissions provide information about cellular properties. Photodetectors convert light signals into electrical pulses that are analyzed. Flow cytometry has various applications including immunophenotyping, cell sorting, DNA content analysis, and cell cycle/proliferation analysis.
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20100617 ps
- 4. Single cell analysis (one at a time)
- 5. Measurements on high number of cells (100 000 cells in less than 2 min, good for statistics)
- 6. Detection of rare events
- 7. …Paulo Salomon 2010.06.17
- 8. Unialgal Clonal Axenic Analysis Isolation forcultivation Freshenvironmentalsamples Service? Traditionalisolationmethods Sorting Coopera-tions BNA Unialgalor Mixed cultures Selectionofcellswith particular traits Analysis Physiology Cultivation Molecular biology Paulo Salomon 2010.06.17
- 9. Whatkindofinformationwegetfrom a flowcytometer? Par value FL1 251 FL2 105 FL3 582 … … SSC 215 FSC 611 EV 265 ToF 590 N cells 35890 Fluorescence Scatteredlight Time ofFlight (ToF) Electronic volume (EV) Cellcounts Paulo Salomon 2010.06.17
- 10. Whatkindofinformationwegetfromtheflowcytometer? Fluorescence Scatteredlight Time ofFlight (ToF) Electronic volume (EV) Cellcounts What can we use this “characterization” for? Paulo Salomon 2010.06.17
- 11. Interpretationoftheflowcytometricsignals Fluorescence Cellularcomposition Physiological status Scatteredlight Celldiameter / lenght Biovolume Internal “granularity” Surface “rugosity” (relationtosizeandform) Time ofFlight (ToF) Electronic volume (EV) Cellcounts Paulo Salomon 2010.06.17
- 12. Fluorescence Cellularcomposition Physiological status Fluorescentdyes Autofluorescence Pigmentcomposition / ratio: Chlorophyll Phycoerythrin Allophycocyanin (red laser) Cellularconstituints: Lipids (NR) Carotenoids (NR) Nucleicacids (SYTO) CH, proteins, … Physiology: Growthphase Stress (Changes in pigmentcomp size, volume, …) Viability/Vitality: Live-deadstains Vitalitystains Metabolism / Stress: Calcein AM (esterase activity) LysoSensor (acidicvacuoles) CSFE (celldivision) … DNA / RNA probes: Typing, detection Contamination: Bacteria (SYTO13) Paulo Salomon 2010.06.17
- 13. Stress Hyperproductivity Maximizationofproductivity Cryopreservation methods Metabolicactivity Cellularcomposition Morphology / complexityandpigments Viability / Vitality FlowCytometric Analysis Pre-screeningforsorting Check forunialgalandmixed cultures Cellcounts Bacteria counts Check foraxenicity Changesover time Growthrates Yield Paulo Salomon 2010.06.17
- 15. Morphology and pigment ratios variability
- 16. Highly relevant internally (bioprospection)
- 18. Morphology
- 20. Metabolism
- 21. Highly relevant internally (bioprospection)
- 22. Highly relevant for the ublic (BNA costumers)Paulo Salomon 2010.06.17