S.varasteh 10/11/2012.

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Presentation transcript:

s.varasteh 10/11/2012

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

Application

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

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

Fluorescence and Fluorochromes

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.

Example of Fluorochromes

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

Using scattered light to determine what is real!

Visualizing Flow Cytometry data

Flow cytometry unstained control

Flow cytometry viability control

Fluorescence Minus One (FMO)

Compensation

Compensation FITC signal is leaking into the PE detector.

Gate on your cell of interest Gating Gate on your cell of interest

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).

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

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.