Flow cytometry works by using a specialized instrument called a flow cytometer to analyze and quantify the properties of individual cells or particles in a sample. The process involves several key steps:
Sample preparation
The sample is first prepared by treating it with fluorescent dyes or antibodies that bind to specific markers on the surface or inside the cell. These markers can be used to identify different cell types or measure other properties of the cells, such as DNA content.
Flow cell
The prepared sample is then loaded into a flow cell, which is a narrow, tube-like channel that allows the cells to flow in a single file stream. This stream of cells is then passed through a laser beam that excites the fluorescent dyes or antibodies, causing them to emit light that is detected by the flow cytometer.
Laser excitation
As the cells pass through the laser beam, they scatter light in different directions depending on their size, shape, and optical properties. The laser beam also excites the fluorescent dyes or antibodies, causing them to emit light that is detected by the flow cytometer.
Optical detectors
The flow cytometer contains multiple optical detectors that measure the light scattered by the cells as well as the fluorescent emissions from the dyes or antibodies. These detectors are able to measure several physical and chemical properties of the cells, including size, shape, granularity, fluorescence intensity, and DNA content.
Data analysis
The data collected by the flow cytometer is then processed and analyzed by specialized software, which can provide detailed information about the properties of the cells or particles in the sample. This information can be used to identify and quantify different cell types, measure cell cycle progression, assess protein expression levels, and analyze cellular signaling pathways.
Overall, flow cytometry is a versatile and powerful technique that allows for the rapid analysis of large numbers of individual cells, providing valuable insights into the complexity of biological systems.
Learn about Sapio Sciences’ flow cytometry data analysis tool here.