The Journal of Clinical and Experimental Pathology (ISSN: 2161-0681) deals with research on infectious disorders associated with immune system and immunological disorders, infectious diseases, treatment of infectious diseases, infectious medicine, epidemiology, diagnostic tests of infectious diseases, infection control, pathophysiology, clinical pathology , preventive medicine. Clinical Pathology deals with patient care, diagnostic services, novel treatments and research on immune infections.
Flow cytometry is a popular cell biology technique that utilizes laser-based technology to count, sort, and profile cells in a heterogeneous fluid mixture. Using a flow cytometer machine, cells or other particles suspended in a liquid stream are passed through a laser light beam in single file fashion, and interaction with the light is measured by an electronic detection apparatus as light scatter and fluorescence intensity. If a fluorescent label, or fluorochrome, is specifically and stoichiometrically bound to a cellular component, the fluorescence intensity will ideally represent the amount of that particular cell component.
Principle of Flow Cytometry
The basic principle of flow cytometry is based on the measurement of light scattered by particles, and the fluorescence observed when these particles are passed in a stream through a laser beam.
- Light scattering results when a particle deflects incident laser light. The extent to which this happens depends on the physical properties of a particle, namely its size and internal complexity.
- Forward-scattered light (FSC) is proportional to the cell-surface area or size of the cell. It is a measurement of mostly diffracted light and detects rays that are just off the axis of the incident laser beam dispersed in the forward direction by a photodiode.
- Side-scattered light (SSC) indicates the cell granularity or internal complexity of the cells. SSC is a measurement of mostly refracted and reflected light that occurs at any interface within the cell where there is a change in the refractive index.
- The measurements of FSC and SSC are used for the differentiation of cell types in a heterogeneous cell population.
- Fluorescent markers used to detect the expression of cellular molecules such as proteins or nucleic acids in a system.
- The fluorescent compound absorbs light energy over a range of wavelengths that is characteristic of that compound.
- This absorption of light causes an electron in the fluorescent compound to be raised to a higher energy level.
- The excited electron quickly decays to its ground state, emitting the excess energy in the form of fluorescence which is then collected by detectors.
- In a mixed population of cells, different fluorochromes can be used to distinguish separate subpopulations.
Parts of Flow Cytometry
A flow cytometer is made up of three main systems: fluidics, optics system, and electronics system.
The purpose of the fluidics system is to transport particles in a fluid stream to the laser beam. The design of the flow chamber allows the sample core to be focused in the center of the sheath fluid where the laser beam then interacts with the particles.
The optical system of the cytometer consists of excitation optics and collection optics. To achieve the specificity of a detector for a particular fluorescent dye, a filter is placed in front of the tubes, which allows only a narrow range of wavelengths to reach the detector.
The electronic system converts the signals from the detectors into digital signals that can be read by a computer. The Analog-to-Digital Converter (ADC) then converts the pulse to a digital number.
On the occasion of its 10 years, Successful Journey, Journal of Clinical and Experimental Pathology decided to provide a partial waiver on its article processing charges to promote quality research from across the nations of the globe to encourage the latest research in the field of Infections, Diseases and Medicine. Journal of Clinical and Experimental Pathology also planning to release a special issue on its new approaches.
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