What is the significance of FSC and SSC in a flow cytometer?

In flow cytometry, the light scattered by cells is measured by two optical detectors: forward scatter (FSC) that detects scatter along the path of the laser, and side scatter (SSC) which measures scatter at a ninety-degree angle relative to the laser.

What is SSC in flow cytometry?

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).

What goes wrong with flow cytometry?

High background/high percentage of positive cells. Two or more cell populations observed when there should be one. High side scatter background (from small particles) Low event rate.

What does SSC mean flow?

side scatter
This parameter is called side scatter (SSC). When measured in conjunction, these two measurements allow for some degree of cellular differentiation within a heterogeneous population. The measurement of forward scatter allows for the discrimination of cells by size.

Why is side scatter SSC directly related to cell complexity?

This is known as Wide Angle Light Scatter (WALS), Orthogonal Light Scatter (OLS), 90° Lights Scatter, or, commonly, Side Scatter (SSC). Side scatter is proportional to cell complexity; the more organelles/bits inside the cytoplasm, the more light scatter, the higher the detected signal.

Why do we use flow cytometry?

Flow cytometry may be used whenever your healthcare provider needs to learn more about the cells inside your body. This type of testing can check the number of immune cells, assess your cell cycle status, identify cancer cells or even analyze your DNA.

What is FACS analysis?

Fluorescence activated cell sorting (FACS) analysis is a derivative of flow cytometry that proceeds in a slightly different direction. The primary objective of FACS flow cytometry is to physically sort a heterogeneous cell sample into separate populations. Isolated cells can then be used for further research.

How do you statistically analyze flow cytometry data?

5 Steps For Accurate Flow Cytometry Statistical Analysis Results

  1. Power the flow cytometry experiment properly.
  2. Establish the threshold (significance level) to your statistical test.
  3. Clearly state the hypothesis.
  4. Choose the correct statistical test.
  5. Know how to plot your data and do it first.

How accurate is flow cytometry?

The diagnostic accuracy of FC was 88.4%, sensitivity was 85.8%, and specificity was 92.9%. In addition, FC accuracy for classes of non-Hodgkin lymphoma was assessed. We conclude that FC is an independently accurate ancillary test in the evaluation of FNA.

What is FSC and SSC?

The light scatter is measured by two optical detectors. One detector measures scatter along the path of the laser (1). This parameter is referred to as forward scatter (FSC). The other detector measures scatter at a ninety degree angle relative to the laser (1). This parameter is called side scatter (SSC).

What does the forward scatter and side scatter in flow cytometry tell about a cell?

Forward versus side scatter (FSC vs SSC) gating is commonly used to identify cells of interest based on size and granularity (complexity). It is often suggested that forward scatter indicates cell size whereas side scatter relates to the complexity or granularity of the cell.