FLOW CYTOMETRYDRM Overview from university
- 1. FLOW CYTOMETRY Dr. MOHAMMED H SAIEMA LDAHR KAAU FACULTY OF APPLIED MEDICAL SCIENCES MEDICAL TECHNOLOGY DEPT. 2ND YEAR MT INSTROMINTATION EXT 21060
- 2. WHAT IS A FLOW CYTOMETER IT IS A MACHINE
- 4. FCM
- 7. WHAT IS FLOW CYTOMETRY ? Flow Cytometry means measuring cells in a fluid medium. It is a technique for making rapid physical and or chemical measurement on particles or cells as they flow in a fluid stream one by one through sensing point . A technology that simultaneously measures multiple characteristics of single cells at a rapid rate. Is the measurement of cellular properties as they move in a fluid past a stationary set of detectors. FCM permits analysis of 105-106 cells per minutes, simultaneously measuring physical and biological properties.
- 8. WHAT IS FLOW CYTOMETRY ? FCM is an automated method used to measure cells or particles as they flow single file though a sensing area. Optical sensing is done with intense light source, usually LASER or mercury arc lamp. The FCM measures light scatter and fluorescent signal generated as cell pass through a light beam.
- 9. WHAT IS THE COMPONANTS OF THE FLOW CYTOMETER ? Instrument Components are, Fluidics Optics Electronics Data Analysis
- 10. Fluidics Cell transportation To introduce and focus the cells for interrogation Most instruments utilize a laminar/sheath flow and turbulent boundary drag techniques to confine (detain) cells to the center of the flow stream. Cells enters the chamber under pressure through a small aperture (opening) which is surrounded by sheath fluid.
- 11. Fluidics The sheath fluid in the sample chamber creates a hydrodynamic focusing effect and draw the sample fluid into a stream. Turbulent boundary flow, The sample is injected into a chamber tapering (narrowing) to a small exit orifice. The speed of the entry of the sample combined with the back pressure generated by the exit orifice results in sample turbulence.
- 13. FACScan FLUIDICS SYSTEM speed of the entry of the sample combined with the back pressure generated by the exit orifice results in sample turbulence.
- 14. Flow Cell Injector Tip Fluorescence Fluorescence signals signals Focused laser Focused laser beam beam Sheath fluid
- 15. OPTICS Excitation optics consist of: Laser: Light Amplification by Stimulated Emission of Radiation Lenses to shape and focus the laser beam Collection optics consist of; i- a collection lens to collect light emitted from the particle-laser beam interaction ii- a system of optical mirrors and filters to route specified wavelengths of the collected light to designated optical detectors
- 16. OPTICS LASER: Light Amplification by Stimulated Emission of Radiation
- 17. OPTICS Laser have the advantage of producing an intense beam of monochromatic light. The most common laser used in FCM are; Argon lasers, produce light between (351-528 nm) Krypton laser, produce light between (350-799 nm) Helium-neon laser produce lines at 543,594,611, and 633nm Helium-cadmium laser produce lines at 325 and 441 nm.
- 18. OPTICS Forward Scatter Channel (FSC): A beam of laser light is projected at specific point of the flow cell . As a particle passes through the laser beam light, most of the light is scattered in the forward direction, this forward scatter is related to the size of the particle.
- 19. OPTICS (FSC) Forward Angle Light Scatter FALS Sensor Laser