The most important technique for the analytical lab is currently flow cytometry (FCM). The most common use of flow cytometry in our lab is for determination of the DNA amount, ploidy level of a plant, to determine aneuploidy, or to detect interspecific hybrids. These assays can be done on leaf tissue, but also directly on the seed.
Flow cytometry is a technique where individual particles are passing in a flow along a measuring point where the particles are illuminated and the scattered light and fluorescence are measured. From a small amount of tissue (leaf or directly from seed) nuclei are isolated. These nuclei are stained with a DNA dye. The flow cytometer analyses the fluorescence intensity per nucleus, which is a measure for the DNA content.
There are two ways of measuring DNA content:
For most measurements, we determine the relative DNA content. This measurement allows for comparison with a control or other samples within the batch. By using an internal standard it's possible to estimate the relative DNA content to within a 1% range. This accuracy allows conclusions to be drawn for many crops about the ploidy level of the plant, to determine Ploidy (PDF), aneuploidy (PDF) or to detect interspecific hybrids (PDF).
We can also measure absolute DNA content. It is sometimes necessary to know the absolute DNA content (in picogram or megabase) for sequencing or other research purposes.
It’s also possible to quantify pollen viability using flow cytometry. Information about the viability of pollen can be used to optimise seed production, storage of pollen, and prediction of seed setting.
Furthermore, Iribov is supplier for flow cytometers and reagents.