The main difference between flow cytometry and FACS is that flow cytometry enables the quick, accurate and easy collection of data on many parameters from a heterogeneous liquid mixture with living cells. But FACS ( fluorescence- activated cell sorting) is a derivative of flow cytometry, which makes it possible to physically sort a heterogeneous mixture of cells into different populations. In addition, flow cytometry uses the different light scattering properties of cells to collect data, while FACS uses highly specific antibodies labeled with fluorescent dyes to differentiate between cell types. In addition, flow cytometry uses a sensor to collect data, while FACS uses an electromagnet to sort the sample.
In short, flow cytometry and FACS are two techniques in analytical cell biology that are used to profile cells in a heterogeneous mixture. In general, flow cytometry involves cell analysis and measurement of protein expression as another analytical technique of FACS that involves sorting out cells in a mixed population.
Key areas covered
1. What is flow cytometry - definition, process, meaning 2. What is FACS - definition, process, meaning 3. What are the similarities between flow cytometry and FACS - overview of the similarities 4. What is the difference between flow cytometry and FACS - comparison of the main differences
Activated fluorescence, FACS, flow cytometry, heterogeneous cell mixing, light scattering
What is flow cytometry?
Flow cytometry is an analytical cell biological technique that allows various parameters of living cells to be determined in a heterogeneous mixture. In addition, the first impedance-based flow cytometry device was invented by Wallace H. Coulter in 1953.
- Cell count
- Determination of cell properties and function
- Detection of microorganisms
- Biomarker recognition
- Evidence of protein engineering
- Diagnosing health disorders such as blood cancer
In addition, the suspension of the sample with multiple cell types in liquid allows it to flow through the flow cytometer. In it, cells ideally flow cell by cell through a laser beam. Here cell types show different light scattering properties that are unique to cell types. This can typically be attributed to the different expression of proteins, nucleic acid content, intracellular components or cell surface components.
In addition, various types of light scattering patterns and fluorescence emission patterns are involved in the analysis of cell types in flow cytometry. They include forward and side scattered light, fluorescence emission, and multiparametric analysis. Forward scattered light from these is refracted by the cell and continues along the same path that it originally traversed. It also helps to know the size of the cell. Meanwhile, laterally scattered light is refracted by the cells in a direction that is outside the original light path. It thus determines the granularity and complexity of different cell types.
In addition, fluorescent molecules from cells emit fluorescent light when excited by the compatible wavelength of the laser, which helps identify different structures of the cells. In addition , fluorescent dyes or fluorescently labeled antibodies can also be used to label specific structures of the cells. Apart from this, forward vs. side scattered light is applied from the leukocyte populations, which aids in the specific identification of granulocytes and lymphocytes.
What is FACS
FACS (fluorescence-activated cell sorting) is a modified form of flow cytometry. The main characteristic of FACS is its ability to physically separate each cell type in the heterogeneous mixture. To do this, this technique uses fluorescently labeled, target-specific antibodies to identify a particular cell type in the mixture. Therefore, FACS separates the heterogeneous cell mixture into two further cell types. Also, FACS was first invented by Len Herzenberg, who later won the Kyoto Prize 2006 for his pioneering work.
In addition, one of the main meanings of cell sorting is to separate cells by phenotype. Thus this enables the analysis of the nucleic acid content, protein expression and metabolic content specific for this phenotype. Second, it helps in the colony formation of healthy and phenotypically defined cells for the generation of stem cells or CRISPR cell lines.
Normally in FACS, the sample flows through a vibrating nozzle that breaks the stream into droplets, which ideally contain a cell. These droplets then pass through an electrical charging ring that physically sorts the cell types according to their charge.
Similarities Between Flow Cytometry and FACS
- Flow cytometry and FACS are two techniques in analytical cell biology.
- In general, they use fluorescence and other properties to detect different cell types in a heterogeneous mixture.
- To do this, they use differences in cell surface components or intracellular components of different cell types.
- In addition, both methods collect forward scatter, side scatter, and fluorescence data .
- In addition, both techniques have applications in molecular biology, genetics, immunology, pathology, and medicine.
Difference Between Flow Cytometry and FACS
Flow cytometry refers to the analysis of biological material by detecting the light absorbing or fluorescent properties of cells or subcellular fractions such as chromosomes when they are passed through a laser beam in a narrow beam. FACS (fluorescence-activated cell sorting) refers to a special type of flow cytometry that provides a method of sorting a heterogeneous mixture of biological cells into two or more containers, one cell at a time, based on the specific light scatter and fluorescence properties of each cell.
Flow cytometry is an analytical cell biological technique, while FACS is a specialized type of flow cytometry.
FACS is followed by flow cytometry, while FACS is the first step in analyzing a heterogeneous mixture of cells.
Flow cytometry involves cell analysis and measurement of protein expression as another analytical technique of FACS, while FACS involves sorting out cells in a mixed population.
Flow cytometry measures the properties of cells such as number, size and nucleic acid content of cells, while FACS separates cells into subpopulations from a heterogeneous mixture.
Flow cytometry uses the different light scattering properties of cells to collect data, but FACS uses highly specific antibodies labeled with fluorescent dyes to differentiate between cell types.
Flow cytometry uses a sensor to collect data, but FACS uses an electromagnet to sort the sample.
In summary, flow cytometry is an analytical cell biological technique to determine the properties of cells in a heterogeneous mixture. Thus, it helps to determine the number of cells, the size and nucleic acid content of the cells, and so on. In addition, different light scattering properties of cells are used, which are unique for each cell type in the mixture. On the other hand, FACS is a type of flow cytometry that enables cells in the heterogeneous mixture to be sorted out into two or more types. It also uses fluorescently labeled antibodies to specifically identify components of different cell types. Therefore, FACS is the first analytical technique, followed by flow cytometry in protein expression assays. Therefore, the main difference between flow cytometry and FACS is the method of cell differentiation and function.
1. Robinson, Ryan, and Stefan Pellenz. “Ryan Robinson and Stefan Pellenz.” Antibodies, December 6, 2013, available here .2. "Flow Cytometry (FCM) / FACS | Fluorescence Activated Cell Sorting (FACS). ”Sino Biological, Sino Biological Inc., Available Here.
1. “Cytometer” by Kierano - Own work ( CC BY 3.0 ) via Commons Wikimedia 2. “Fluorescence-assisted cell sorting (FACS) B” by SariSabban ( CC BY-SA 3.0 ) via Commons Wikimedia