1. s.varasteh
10/11/2012

2. اصول و کاربرد های فلوسایتومتری تحقیقاتی
اصول و کاربرد های فلوسایتومتری تحقیقاتی
دانشگاه علوم پزشکی وخدمات بهداشتی-درمانی استان قزوین

4. Application

5. What is Flow Cytometry?
The measurement of cell in flow stream which delivers the cells in single file past a point of measurement.

9. Flow cytometers are engineered with precise light pathway.
Light is routed through three different types of filter.

10. Fluorescence and Fluorochromes

11. Fluorochromes
Fluorochromes absorb energy (light) and their electrons go from a ground state to an exited state. The electrons return to ground state by emitting light of lower energy , therefore longer wavelength.

12. Example of Fluorochromes

13. The fluorescence intensity measured is proportional to the number to the fluorescent molecules bound to the cell.

20. Using scattered light to determine what is real!

22. Visualizing Flow Cytometry data

23. Flow cytometry unstained control

24. Flow cytometry viability control

26. Fluorescence Minus One (FMO)

28. Compensation

30. Compensation
FITC signal is leaking into the PE detector.

31. Gate on your cell of interest
Gating
Gate on your cell of interest

32. When there is an obvious positive and negative population it is easy to use a control (A) to set a marker to define positivity (B).

33. Control
Negative sample (A) and test sample (B) and  (C) shows that probably all cells are weakly positive.

34. Negative sample (A) and test sample (B)
Negative sample (A) and test sample (B). The overlay (C) shows that some cells are negative and some are positive.