![flowjo 10 restrict cell number in dot plot flowjo 10 restrict cell number in dot plot](https://www.bdbiosciences.com/content/dam/bdb/products/global/reagents/flow-cytometry-reagents/research-reagents/single-color-antibodies-ruo/564034_base/564034Image1.png)
Note: This example is for illustration purposes only, the populations illustrated are either not actual monocyte/lymphocyte cells, or the patient is exhibiting acute monocytosis. Discrimination of lymphocytes and monocytes in FlowJo ™ based on scatter parameters.
![flowjo 10 restrict cell number in dot plot flowjo 10 restrict cell number in dot plot](https://docs.flowjo.com/wp-content/uploads/sites/6/2013/06/11ColorSmall.fcs_-Ungated-FlowJo-3-1.png)
In general, monocytes are larger than lymphocytes and exhibit forward scatter of a higher intensity.įigure 2. Monocytes and lymphocytes are two classes of white blood cells. The intensity of the produced voltage is proportional to the diameter of the interrogated cell (1).įSC is helpful for distinguishing between cells of the immune system. Forward scatter is detected by a photodiode, which converts the light into an electrical signal. FSC intensity is proportional to the diameter of the cell, and is primarily due to light diffraction around the cell. The measurement of forward scatter allows for the discrimination of cells by size. When measured in conjunction, these two measurements allow for some degree of cellular differentiation within a heterogeneous population. This parameter is called side scatter (SSC). The other detector measures scatter at a ninety degree angle relative to the laser (1). This parameter is referred to as forward scatter (FSC). One detector measures scatter along the path of the laser (1). The light scatter is measured by two optical detectors. A wavelength of 488 or 405 nm is common (1). Accordingly, the laser usually emits light at a wavelength shorter than interrogated particles. When cell size is smaller than the excitation of the interrogating laser, the scatter behavior is inconsistent and low-intensity. The ratio between cell size and laser wavelength alters scatter behavior. Figure 1 provides an overview of a flow cytometer schematic.įigure 1. While being interrogated by the lasers, the cell scatters light (1). The population then flows past a set of laser light sources one cell at a time. The suspension is funneled through a nozzle that forges a single-cell stream. In a flow cytometer, a cell population is suspended in a clear saline solution. Flow cytometry is a method of single-cell analysis that includes the characterization of a cell's physical properties.