How do you analyze flow cytometry results?

Flow cytometers utilize properties of fluid dynamics to send cells one at a time through a laser. The optics and computer systems then track the photon emission from excited cells and analyze both the light that scatters past (forward scatter; FSC) and the light that scatters perpendicularly (side scatter; SSC).

What does a flow cytometry histogram show?

Flow cytometry histograms are a direct tabulation of the frequencies of measured values in a fixed number of channels or bins. They are often described as being displayed in a particular scale without reference to the underlying process of binning.

What happens when you increase the flow rate on a flow cytometer?

(A) With traditional hydrodynamic focusing, increasing the flow rate (and pressure) allows more than one cell at a time to pass through the interrogation point.

What is the effect of increasing the flow rate of the sample when carrying out flow cytometry to Analyse a cell population?

Increasing the sample pressure increases the flow rate by increasing the width of the sample core. This, in turn, allows more cells to enter the stream within a given moment. With a wider sample core, some cells could pass through the laser beam off-center and intercept the laser beam at a less optimal angle.

How do I know if my flow cytometry data is bad?

How to Identify Problems with Flow Cytometry Experiment Design – Bad Data Part 3

1. Experiment lacks single stain controls.
2. Only compensation beads run for single stained controls (no single stained cells).
3. Use of isotype controls instead of FMOs.
4. Unlabeled parameters and/or tubes.

What is flow cytometry Slideshare?

• Flow cytometry is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles • A sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument.

How is fluorescence activated for FACS?

An antibody specific for a particular cell surface protein is associated to a fluorescent molecule and then added to a mixture of cells. Droplets containing a single cell are then detected by an electric field into collection tubes according to their charge. …

What happens in flow cytometry?

Flow Cytometry is a technique used to detect and measure physical and chemical characteristics of a population of cells or particles. In this process, a sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument.

What is gain in flow cytometry?

Two types of photon detectors are used to amplify and then convert light into electronic signals in flow cytometry. Increasing the voltage increases the energy of the electrons and thereby amplifies the signal. A measure of the amplification is a unitless quantity, called the Gain. In a PMT, the gain isn’t linear.

What does SSC measure?

Side-scattered light (SSC) is proportional to cell granularity or internal complexity. SSC is a measurement of mostly refracted and reflected light that occurs at any interface within the cell where there is a change in refractive index (Figure 3-1).