Applications of Flow Cytometry

Flow Cytometry is a technique of identifying and measuring the physical and chemical properties of cells which travel in a suspension. The benefit of this technique is that it allows the rapid and simultaneous measurement of several parameters on a cell by cell basis. It can detect and analyze thousands of cells per second.

There are wide range of Applications of flow cytometry, some of them are explained below.

1) Immunological Applications:

Immunophenotyping (It involves identification and characterization of cell surface proteins (CD markers) or intracellular proteins using fluorescent-conjugated antibodies. For example: identification of CD markers present in blood cells.)

Stem Cells Analysis ( involves studying of stem cells such embryonic stem cells (ESCs), human induced pluripotent stem cells (hiPSCs), studying the pluripotency and multipotency. Characterization of multiple antigens present on the cell surface or intracellular using multi-color flow cytometry methods).

Cell Counting

Cytokine Analysis and Bead-Based immuno assays.

Platelets Identification.

2) Cell or Cancer Biology Applications:
Apoptosis Studies

Cell Cycle Analysis

DNA Damage

Cell Proliferation Studies.

Cancer Research

Cell Signalling Studying

Calcium Flux Studies

Gene Expression Analysis

Kinetics Studies

3) Microbiology Applications:
Microbial Viability

Microbe Counting

Absolute Counting of Microorganisms

Water Quality Analysis

Aquatic Research

Microbe Species Identification

Microbe Species Characterization

4) Flow Cytometry also has applications in Industry level such as in Bio-processing, Bio-fuels, Plant Biology, Foods and Beverages.

5) It has application in Veterinary Also., such as identification and separating sperm based on X- and Y- chromosomes.

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Statistics in Flow Cytometry Data and "MFI" values

The speed of the flow cytometry offers wide range of data points and data plots. Due to its sensitivity and versatility, it has been used extensively. Flow cytometer can detect up to 1000s of cells per second.. But, it should be noted that the flow cytometry also involves the statistics, its significance, calculating the fluorescence intensity.. Statistics in flow cytometry involves, total number of data points acquired, percentage population, most importantly mean and median fluorescence intensity, and others. Most of us get confused about the term "MFI", whether it means mean fluorescence intensity or median fluorescence intensity.. Truth is, both.. It means it can be described as either mean or median fluorescence intensity. Some researchers use Mean for MFI and some use Median for MFI values. (Keep...

Nanodrop

Thermofisher, Nanodrop 1000 Nanodrop is a spectrophotometer which is designed to measure the concentration of nucleic acids at 260nm and Proteins at 280nm present in a very small minute volume. It is also capable of measuring the concentrations of dsDNA, RNA, ssRNA and purified protein. Required sample size is usually 1µL. Nanodrop instruments are usually full spectrum spectrophotometer, i.e., Ultra violet and visible range wavelengths (250nm to 750nm). Nanodrop is commonly used for the quantification of the DNA concentration and RNA concentration in a given sample. However there are many other applications which can be performed on this instrument. It works on the principle of Spectrophotometer or Sample Retention System.... Principle of Nanodrop: "The Nanodrop works on the principle of sample retention system.. When the small volume of sample is introduced, the arm and pedestal will work together and make the sample in the form of colum...

Compensation in Flow Cytometry (Fluorescence Spill Over Correction).

The term "Compensation" in flow cytometers refers to the process of correcting fluorescence spill over. The compensation matrix ensures that the fluorescence emitted is detected in a particular detector which is from the fluorescence being measured. As we all know that most of the fluorescence molecules have broad emission range. if more than one one fluorescence is selected, their emission spectra may overlap. To correct this spectral overlap in cytometry data, a mathematical matrix called compensation is used. The Intention of this article is to teach you the compensation from basic theory to practical. This example for FITC and PE fluorescence molecules shows that, Some of the FITC fluorescence is being detected in PE Detector. and Some of the PE fluorescence is being detected in FITC Detectors. Task here is to make sure that in cytometry data, FITC is in FITC detector only and PE is in PE Detector only....

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