Federal Institute for Materials Research and Testing (BAM), Biophotonics Division 1.2, Richard-Willstätter-Str. 11, D-12489, Berlin, Germany.
Department of Physics, Humboldt-Universität zu Berlin, Newtonstr. 15, D-12489, Berlin, Germany.
Sci Rep. 2018 Nov 13;8(1):16715. doi: 10.1038/s41598-018-35137-5.
Time-resolved flow cytometry represents an alternative to commonly applied spectral or intensity multiplexing in bioanalytics. At present, the vast majority of the reports on this topic focuses on phase-domain techniques and specific applications. In this report, we present a flow cytometry platform with time-resolved detection based on a compact setup and straightforward time-domain measurements utilizing lifetime-encoded beads with lifetimes in the nanosecond range. We provide general assessment of time-domain flow cytometry and discuss the concept of this platform to address achievable resolution limits, data analysis, and requirements on suitable encoding dyes. Experimental data are complemented by numerical calculations on photon count numbers and impact of noise and measurement time on the obtained lifetime values.
时间分辨流式细胞术是生物分析中常用的光谱或强度多重检测的一种替代方法。目前，关于这个主题的绝大多数报告都集中在相位域技术和特定应用上。在本报告中，我们提出了一种基于紧凑设置和利用纳秒级寿命编码珠进行简单的时域测量的时间分辨检测的流式细胞术平台。我们对时域流式细胞术进行了全面评估，并讨论了该平台的概念，以解决可实现的分辨率限制、数据分析以及对合适编码染料的要求。实验数据通过对光子计数数的数值计算以及噪声和测量时间对获得的寿命值的影响得到补充。
