What is FSC and SSC in flow cytometry?

Answer. 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 the difference between FACS and flow cytometry?

FACS is used as a cell sorter and enriched for a subset of cells which is often then studied in further detail using flow cytometry or other analytical techniques2. Flow cytometry is used for cell analysis and is focused on measuring protein expression or co-expression within a mixed population of cells.

What is the basic principle and application of FACS?

The basic principle of flow cytometry is the passage of cells in single file in front of a laser so they can be detected, counted and sorted. Cell components are fluorescently labelled and then excited by the laser to emit light at varying wavelengths.

What is SSC A and 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).

What is FSC a FSC H FSC W?

The purpose of both FSC-H and FSC-W is to check the doublet or multiplet cells, based on FACS machine people generally use FSC-A vs FSS-W or FSC-A vs FSC-H to exclude multiplet. It is recommended that if FSC-A vs FSC-H is used then for better resolution of multiplet one should also use SSC-A vs SSC-W/H.

Does FACS Count cells?

yes its better. The antibody you use should be titrated on a specific number of cells. If you deviate too much from that number you will end up with an insufficient amount of antibody to properly stain your cells. You will underestimate the number of true positive in your samples.

What are the three main systems in a flow cytometer and its function?

A flow cytometer is made up of three main systems: fluidics, optics, and electronics. The fluidics system transports particles in a stream to the laser beam for interrogation.

What is the main principle of flow cytometry?

Flow cytometry (FCM) is a technique which enables rapid analysis of statistically significant number of cells at single cell level. The main principle of this technique is based on scattering of light and emission of fluorescence which occur when a laser beam hits the cells moving in a directed fluid stream.