Why use flow cytometry for DNA analysis?
Determining genome size and ploidy is a particularly important form of analysis in aquaculture, helping breeders select the specimens having the best properties. It also plays a role in basic research in evolutionary biology, taxonomy and ecology. The traditional method of counting chromosomes by classical light microscopy, assessing nuclear volume or karyotyping, is both time-consuming and imprecise. Flow cytometry is the state-of-the-art method to accurately measure ploidy and genome size: it is elegantly performed in four simple steps, including in the end the analysis of hundreds of nuclei per second.
“Flow cytometry is the state-of-the-art method in the breeding industry and in research for determining both
the ploidy level and genome size in plants, animals and microorganisms.”
While the primary use of flow cytometry in aquaculture was initially for DNA analysis, its use has been naturally extended to analysis of fresh or marine water habitat. Application areas include:
- Animal ploidy control
- Genome size and C-value determination
- Aneuploidy analysis
- Hybrid detection
- Enumeration of micro-organisms in water
- Viability of water microbes
Sysmex DNA analysis solutions
Based on the proof of concept of its predecessors, Sysmex is proud to offer you up-to-date flow cytometry solutions for determining nuclear DNA that are designed specifically for this application.
From staining reagents and control cells to our third-generation flow cytometry analysers – CyFlow Ploidy Analyser and CyFlow Space DNA analysis configurations – Sysmex DNA analysis solutions are designed to be simple, accurate and fast.
Efficient and validated workflow
Samples are stained in four simple steps:
- mechanical release of nuclei using gentle extraction reagents
- filtration through CellTrics filter
- addition of a ready-to-use staining reagent and
- automated analysis of hundreds of nuclei per second.
Obtain precise results
The unique configurations of our DNA analysers, optimised for measurement of propidium iodide (PI), 4',6-diamidino-2-phenylindole (DAPI), or both, provide superior results: A 532 nm laser combined with PI provides superior results compared to a standard 488 nm laser for analysis of genome size. Our unique UV LED (365 nm), optimal for measurement of DAPI, offers the high resolution required for ploidy analysis.
Control cell reagents are well suited to check resolution and linearity since the same cells are present not only as single cells but also as aggregates of two, three and more cells.
Obtain results quickly
Many aspects of our analysis system work together to speed up analysis, providing ploidy and genome size results in less than two minutes:
- reduced acquisition time per sample
- autoloader capability allows analysis of a 96-well-plate within 15 minutes
- fast statistical analysis with the option to define peaks automatically
- peak analysis function produces analytical ranges automatically
- minimal analyser cleaning times