Flow cytometry is a widely used technique in drug discovery, genomics, immunoprofiling for analyzing the expression of surface protein and intracellular molecules, characterizing and sorting cell populations, assessing the purity of isolated subpopulations, and cell-based assays such as proliferation, cyto toxicity and cell death.
A cell suspension is passed through a flow chamber single file, through one to several lasers to detect the scattered light and fluorescence of single cells. By using fluorescently-labeled antibodies, a flow cytometer is capable of detecting and differentiating between multiple proteins of interest. When used multi-parametrically, this technique forms a “panel,” which is defined by the multiple fluorochromes spanning the light spectrum, each tagged to an analytically relevant protein target.
Conventional flow cytometers have difficulty distinguishing fluorochromes with highly overlapping peak emissions, severely limiting fluorochrome choice and multiplexing capability, leading to misinterpretation of the result.
Using FSPTM where entire emission spectrum is captured across the different modules and then stitched together to create a spectral signature that combines emission information from all excitation wavelengths , Cytek systems can detect the entire fluorochrome emission, allowing researchers to use highly overlapping dyes together, to extract autofluorescence,to run panels of up to 40 colors and beyond without sacrificing data resolution.
No longer limited by the conventional peak emission bandpass filter paradigm, researchers can now visualize the full spectrum characteristics of each fluorochrome across each laser line, look for new spectrally unique reagents to integrate into continuously expanding panel designs and get more results in one experiment.
Cytek’s experience in designing highly multiparametric panels has led to a deeper understanding of how fluorochromes react together in increasingly complex fluorescent experiments. Using this expertise, Cytek developed and launched the new cFluor reagents, optimized for use in full spectrum panels.
The optical design combined with the unmixing capability in SpectroFlo software allows you to easily expand panels. The 3 laser configuration provides outstanding multiparametric data for a wide array of applications.
Expanding to 20 Colors: A 20-color panel preserving the original 7-color assay is summarized in the table below. Reagents shared by the 7-color and 20-color panels are shown in bold.
NanoFCM provides a versatile and powerful platform — Flow NanoAnalyzer for the multiparameter analysis of individual extracellular vesicles (30-150 nm in diameter), the fluorescence of single phycoerythrin molecules can be well detected against the background. The Flow NanoAnalyzer platform enables quantitative and multiparameter analysis of single EVs down to 40 nm, which is distinctively sensitive, yet high-throughput, and shows great potential in liquid biopsy applications.
Flow NanoAnalyzer is expected to become a powerful tool for life science, nanoscience and nanotechnology studies. Flow NanoAnalyzer can be used for the multiparameter characterization of natural and synthetic nanoparticles (7-1000 nm) at the single-particle level, such as extracellular vesicles, mitochondria, bacteria, viruses, nanomedicine and naonomaterial. Combining light scattering and fluorescence detection, high-resolution distributions of particle size and biochemical properties can be acquired simultaneously in 1-2 minutes.
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