What is flow cytometry?

Flow cytometry (FCM) is a method for the measurement of optical properties (fluorescence, light scatter) of isolated particles (cells, nuclei, chromosomes) that move one by one in a narrow liquid stream through a powerful beam of light.

Currently, FCM is routinely used in clinical diagnostics, biotechnology, and basic and applied research (immunology, molecular biology, genetics, pharmacology, zoology, marine biology, and botany).

It facilitates determination of various characteristics at both cellular (size, shape, granularity, membrane potential, cell cycle, apoptosis) and intracellular levels (DNA and RNA contents, base composition, protein content, intracellular pH, calcium concentration, chromosome size, centromeric index, etc.).

Rapid estimation of nuclear genome size is the most frequent cytometric application in plant biosystematics.

Advantages of FCM

• rapid, easy and convenient sample preparation and analysis (allows investigation of several dozens or even hundreds samples per working day)
• high accuracy (facilitates detection of minute variation in genome size)
• investigation of mitotically inactive cells from a broad variety of plant tissues (leaves, stems, roots, sepals, petals, seeds)
• non-destructiveness (opens way for comprehensive investigation of rare and endangered species or seedlings in very early ontogenetic stage)
• rapid detection of mixed samples or endopolyploidy (records separately the characteristics for each object)
• low operating costs

Limitations of FCM

• general requirement of fresh plant material
• problematic detection of aneuploidy or accessory chromosomes
• difficult analysis of species with high level of secondary metabolites (tannins, mucilaginous compounds, etc.)