Since the first commercial flow cytometers capable of measuring a single fluorescence parameter were introduced in the early 1970s, the technology has greatly evolved. Continued development of flow cytometry technology resulted in an ever-increasing number of lasers and detectors, allowing detection of greater numbers of parameters.
However, the opportunity for more persisted. Researchers were interested in measurement across the complete fluorescence spectrum. Ultimately, in 2012, the first commercial spectral flow cytometer was released.
In conventional flow cytometry, each fluorophore present is measured in a single target detector with a portion of the full emission collected using band-pass or long-pass optical filters. Spillover from other fluorophores that may have emission in that detector is corrected using compensation.
In contrast, spectral flow cytometry uses multiple detectors to measure the full spectrum emission of every fluorophore across multiple lasers used in the system to create a more detailed signature for each fluorophore. The spectrum detected by each group or array forms a spectral signature.
While conventional flow cytometry uses compensation to correct for fluorescence spillover, spectral flow cytometry uses a process called unmixing to identify each fluorophore. Spectral unmixing uses a mathematical algorithm that distinguishes the many fluorophore signatures within a multicolor sample, based on the unique spectral signature of each fluorophore. Through this approach, fluorophores with near-identical peak emissions but different off-peak emissions may be distinguished and used together in a panel.
For example, detecting rare or low-abundance cell populations can be challenging with conventional cytometers. Spectral flow cytometers—such as Thermo Fisher Scientific’s Attune Xenith—improve the detection of rare events and subtle differences within cell populations. The identification and analysis of rare cell populations, such as stem cells or circulating tumor cells, is crucial for applications requiring high precision and accuracy.
Accurate and comprehensive multicolor analysis
The Attune Xenith Spectral Flow Cytometer's high sensitivity, flexibility and advanced spectral capabilities make it suitable for handling challenging sample types without clogging—resulting in smooth sample acquisition and reliable data analysis.
With expanded optical capabilities, including 6 lasers, 51 fluorescent detectors and 6 scatter channels, the Attune Xenith Flow Cytometer supports both traditional compensation and spectral unmixing analysis. It is also compatible with Invitrogen CytKick autosamplers for efficient handling of 96- and 384-well plates.
Researchers often need to adapt their experiments to various applications and sample types. The Attune Xenith offers flexible configurations and compatibility with a wide array of fluorophores and reagents. Additionally, the acoustic focusing system reduces the need for frequent maintenance and calibration, minimizing downtime and operational costs. The instrument features an intuitive software interface and automated setup procedures, making it accessible to users with varying levels of flow cytometry experience. This reduces the learning curve and enhances productivity in the lab.
Lastly, the Attune Xenith is equipped with advanced software tools that facilitate efficient data acquisition, analysis, and visualization, streamlining the workflow for researchers.
Collectively, these features and more make the Attune Xenith Flow Cytometer a versatile tool that addresses many of the limitations of traditional flow cytometers, providing researchers with enhanced capabilities and maximized productivity.