What is forward scatter in flow cytometry?
One detector measures scatter along the path of the laser (1). This parameter is referred to as forward scatter (FSC). The measurement of forward scatter allows for the discrimination of cells by size. FSC intensity is proportional to the diameter of the cell, and is primarily due to light diffraction around the cell.
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 is FSC and SSC in FACS?
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 are the two types of scattering used to analyze cells in flow cytometry?
Scattered Light Because of the nature of its collection, this parameter is referred to as Small Angle Light Scatter (SALS), Forward Angle Lights Scatter (FALS), or, most commonly, Forward Scatter (FSC).
How does 7aad stain dead cells?
Several different fluorochromes can be used to stain non-viable cells including 7-amino actinomycin D (7-AAD). 7-AAD is a membrane impermeant dye that is generally excluded from viable cells. It binds to double stranded DNA by intercalating between base pairs in G-C-rich regions.
What is PE flow cytometry?
PE & APC. Phycoerythrin (PE) and allophycocyanin (APC) are members of a protein family called phycobiliproteins. This family of proteins is present in photosynthetic organisms, such as algae and cyanobacteria. For use in flow cytometry, PE and APC are extracted from red algae and purified.
What is FSC A?
forward scatter area (FSC-A) density plot can be used to exclude doublets as shown in Figure 3 below. A side scatter height (SSC-H) vs side scatter area (SSC-A) plot can also be used. This type of gating can be more sensitive for doublet exclusion as the FSC detector is not usually a photomultiplier tube (PMT).
Is 7-AAD toxic to cells?
I didn’t observe any toxicity due to 7AAD in the samples and results are just fine everytime. I will recommend using them together to avoid the over complications in protocol. 7AAD binds well in Annexin binding buffer also. You can wash the dyes off by PBS once incubation is completed.
What is forward scatter?
In physics, telecommunications, and astronomy, forward scatter is the deflection—by diffraction , nonhomogeneous refraction, or nonspecular reflection by particulate matter of dimensions that are large with respect to the wavelength in question but small with respect to the beam diameter—of a portion of an incident electromagnetic wave, in such a
What do the results of my flow cytometry mean?
What Do the Results Mean? Flow cytometry can provide information that is used to diagnose, stage and monitor blood cancers. It can also be used to test for minimal residual disease (MRD), the number of cancer cells remaining in the body after treatment.
What is FSC and SSC in flow cytometry?
Using Flow Cytometry one can determine relative size of cells or events using a known control. Using the FSC and SSC parameters one can look at a mix of cells and distinguish them from one another based off of size and internal complexity. Along the X-axis is the FSC(Forware SCatter) parameter.
What are the advantages of flow cytometry?
The two greatest advantages of flow cytometry are its ability to measure a large number of parameters (2 to 30 or more) on the same sample and its ability to collect information from millions of cells in a matter of seconds.