Evaluating spectral cytometry for immune profiling in viral disease

Few comparisons have been reported between conventional and spectral cytometry systems. In a recent issue of Cytometry Part A, Niewold et al. conducted such a comparison and assessed the quality of the resulting datasets. Signal intensity, spread, and resolution were compared along with different methods of separating fluorophore signals. Within the spectral data, signal spread and resolution were comparable using compensation and unmixing methods. However, for some highly overlapping fluorophores, unmixing resolved the two fluorescence signals where compensation did not. Data from mid‐ to large‐size panels were acquired and were found to have comparable resolution for many fluorophores on both instruments, but the spectral system had improved signal resolution for several fluorophores. Furthermore, autofluorescence extraction on the spectral system allowed for greater population resolution in highly autofluorescent samples. Overall, the implementation of a spectral cytometry approach yielded data comparable to that generated on conventional systems, with several potential advantages afforded by the larger number of detectors, and the integration of the spectral unmixing approach.

Paula Niewold et al. Cytometry Part A, 2020 DOI: 10.1002/cyto.a.24211

评估光谱细胞术对病毒性疾病的免疫分析

常规和光谱细胞计数系统之间的比较报道很少。在最近的《细胞计数法》 A部分中，Niewold等。进行了这样的比较并评估了所得数据集的质量。比较了信号强度，扩展和分辨率以及分离荧光团信号的不同方法。在光谱数据内，使用补偿和解混方法可比较信号扩展和分辨率。但是，对于某些高度重叠的荧光团，通过解混可以分辨出两个荧光信号，而补偿信号却没有。采集了中大型面板的数据，发现两种仪器上许多荧光团的分辨率均相当，但是光谱系统对几种荧光团的信号分辨率有所提高。此外，在光谱系统上的自发荧光提取可以在高度自发荧光的样品中实现更大的种群分辨率。总体，

Paula Niewold等。细胞计数法A部分，2020 DOI：10.1002 / cyto.a.24211