What is the limitations of flow cytometry?

There are a number of both limitations and advantages to using flow cytometry and fluorescent automatic cell sorting. Limitations are that the machines are costly, and the training needed to properly use them is extensive. The main advantage is that they reduce considerably the time cost associated with isolation work.

What is the limit of sensitivity for a typical flow cytometer?

Most cytometers can detect cells between 1 and 15 microns in diameter, although it is possible to detect particles outside of this range (0.2 -150 microns) using specialized systems [1].

What is the sensitivity of flow cytometry?

Second, a flow cytometric protocol for staining and acquisition of large numbers of cells (>4 million) was developed, allowing theoretical sensitivities of at least 0.001\% (≤10−5).

What can you use instead of flow cytometry?

All Answers (7) Hi Jerome, alternative methods could be 1) an immunofluorescent microscopy or 2) a CD4/CD8 specific isolation and counting. 1) For microscopy, you will need of course corresponding microscope: you have to fix and stain your cells in a plate with mentioned primary antibodies (GK1.

What is the 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.

Is limit of detection sensitivity?

Detection limit is a measure of the smallest concentration which can be determined with a specified precision or reproducibility. As defined, the detection limit is a function of both signal strength (or sensitivity) and signal stability.

Why is pressure so important for flow cytometry?

The pressure of the sheath fluid sets the speed of the system, so if you want to change the event rate—that is, the number of cells or particles passing the interrogation point in a given period—you will have to change the differential pressure between the sample and sheath fluid.

What is the number of events per second flow cytometry?

Best practice for rare event analysis is to run the system at low differential pressure so that the event rate is no more than 10,000 events per second (depending on your instrument). It is often even better to run at a lower rate, such as 5,000 events per second.

What advantage do flow cytometry has over immune florescence microscopy?

Flow cytometry, however, can handle larger volumes of up to hundreds of thousands of cells. Furthermore, flow cytometry can use fluorescent properties to distinguish and sort living cells in the thousands, while fluorescence microscopy is incapable of this.

Is flow cytometry A FACS?

FACS is an abbreviation for fluorescence-activated single cell sorting, which is a flow cytometry technique that further adds a degree of functionality. The technology to physically sort a heterogeneous mixture of cells into different populations is useful for many therapeutic and clinical applications.

What is the principal disadvantage of flow cytometry methods for cell death?

The principal disadvantage of flow cytometry methods for cell death is over interpretation of the data. What the flow cytometer tells us is the amount of apop-tosis that is detectable at the time of sampling.

What is a flow cytometer used to measure?

However, the flow cytometer is also well suited to analyzing cell death, and techniques have capitalized on the biological characteristics of apoptosis and necrosis to identify and quantify cell death and to distinguish between the two forms.

What does a flow cytometer tell you about apoptosis?

What the flow cytometer tells us is the amount of apop-tosis that is detectable at the time of sampling. It tells us nothing about the rate of apoptosis nor anything about the total amount of cell death that may be present in the cell population as a whole.