The basic principle of flow cytometry is to determine the cell characteristics based on the movement of the particles. The particle movement is due to how cells are suspended. Cytometry is the analysis of cellular components and cellular processes. Flow cytometry is a specific technique used to examine the composition and functionality of living cells. Cytometry can explore a wide variety of biological processes in vitro. The field of cytometry as a whole is known as flow cytometry. Luckily for you, you don’t need to be a cytometry expert in using flow cytometry in your lab. The primary function of flow cytometry is to detect and analyze different cell properties. The technique is used to determine the characteristics of red blood cells, white blood cells, platelets, and DNA and RNA. The fields that use this method are abundant and varied.
- What Is Flow Cytometry?
Flow cytometry is a method used to examine the composition and functionality of living cells. In inflow cytometry, cells are treated with chemicals that make them fluoresce or alter their shape, allowing them to be measured. This is called flow cytometry. This measurement can be used to determine cell characteristics such as cell number and size, cellular function (such as movement), and cellular structure (such as membrane properties).
- Fluid Dynamics
The first principle of flow cytometry is fluid dynamics. Fluids in motion are easier to observe. For example, blood cells can be regarded as flowing through a tube. The liquid in the tube will move at different speeds depending on what is in it and where it is going. The speed at which the blood flows through the pipe will depend on how much force is pushing the blood through, such as gravity or pressure from a pump. If there is no force pushing blood through the tube, it will take longer for the fluid to reach its destination. Because of hydrodynamic focusing, when the heterogeneous sample is placed in the channel, the cells are moved through the track at a lower speed. This means that they move through the channel slower than other cells to be observed more easily.
- Optics
Optical microscopy is the technology that is used to observe the cells. The microscope will contain a lens focused on the sample and a light source to illuminate the sample. The microscope will have a stage to move the cell in different directions and positions. This allows for optical backscatter, which is when light emitted by a cell enters an optical channel but does not go out of it. This observation method increases the chances of observing cells after they are treated with chemicals or fluorescing. The amount of light that is incident on the cells will depend on the light source used. For example, the white light will be more intense than red light and thus more easily observed. The type of microscope used also affects how well the cells can be detected.
- Electronics
The electronics used in flow cytometry are essential to the process. The electronics contain a computer that processes the data and a “detector” to measure the cells. The computer can be programmed to make it easier to analyze the data, such as focusing on specific cells or groups of cells. It also allows for predicting which cell properties will be most prominent, such as membrane permeability or changes in surface charge. The detector can detect various things measured from the cells, such as fluorescence intensity, size, and shape. The sensor can also calculate how many cells are present in the sample at any given time. This ability is called a “flow rate.” The electronics also control the channels through which the cells are passed. These channels are needed to measure cell characteristics such as size, shape, and movement. Without them, cells would not be able to move through the channel at all.
Flow cytometry is an essential part of cell biology. Flow cytometry allows us to observe cells more efficiently and accurately than ever before. The amount of information obtained from one sample is enormous, and the ability to analyze different types of cells gives researchers a way to study many various aspects of cells.
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