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Flow cytometry

An explanation of flow cytometry or FACS (fluorescence-activated cell sorting) and how it's used in cell sorting, immunophenotyping, and other applications.


Factsheet

Flow cytometry or FACS (fluorescence-activated cell sorting) refers to the usage of specialised instruments that measure and characterise cell populations.

Characterisation is based on the physical properties of the cells (size, volume, internal complexity/granularity) and/or on individual or combination of proteins that are specifically localized in distinct sub-populations. These proteins are stained with highly specific fluorescent markers like in fluorescence microscopy.

One can either analyse the composition of a mixed cell populations and extract statistical information about each sub-population, or physically collect pure sub-populations of interest in an individual container (tube or well-plate) in what is called cell sorting.

In principle, FACS drives individual cells through a laser beam, and the scattered light and/or the fluorescence emitted is collected and analysed by an array of detectors, collecting various data for each cell scanned. Hundreds or thousands of cells per second can be analysed at any single time, making flow cytometry a reliable and fast technique to extract variable information from thousands of cells in the matter of minutes.

Application of flow cytometry include: cell sorting, immunophenotyping, cell cyle analysis, apoptosis, cell proliferation assays, cell signaling, intracellylar calcium flux.

Flow cytometry equipment at the University

BD FACSAria III cell sorter

The FACSAria is an advanced four-way cell sorter offering high-speed multicolour analysis and cell sorting. It has four lasers (407/488/561/633) and can analyse up to 13 fluorescent channels.

The system supports four nozzle diameters (70, 85, 100, and 130μm) enabling the sorting of a wide range of particle sizes, from bacteria to large mammalian cells.

The sort collection chamber allows two-way or four-way sorting into tubes while the automatic cell deposition unit (ACDU) allows sorting into slides or multi-well plates as well as index sorting. ​

Cell sorting is ideal for recovering specific cell subpopulations for reculturing, selective cloning, enrichment, and purification before Polymerase Chain Reaction (PCR) amplification or protein analysis.

Sony ID7000 full spectrum cell analyser

The ID7000 is a latest-generation full spectrum cell analyser (flow cytometer) – a technology that doesn’t rely on filters and gives great flexibility on panel design. Our system is equipped with five lasers, (355/405/488/561/637nm) and, under the right conditions, you could analyse more than 40 colours in a single sample.

The system's sensitivity is superb, making detection and analysis of dim or rare populations easier, while its automated tasks can reduce the effort required to analyse multiple samples. The system supports any kind of 96-well plate, flat-bottom 384-well plates, and 5ml tube racks (24 tubes per rack).​

Features like the Spectral Library and the Autofluorescence Finder simplify setup and increase the accuracy of the data you acquire. Intuitive tools and guided workflows make using it straightforward.​

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