Appl Opt. 2022 Jun 10;61(17):5067-5075. doi: 10.1364/AO.458807.
A 64-channel detection system for laser-induced fluorescence (LIF) detection at the cell level is established and applied to single event counting. Generally, fluorescence detection at the cellular level requires a dyeing label to enhance the scattered light intensity for the photodetector. However, the dyeing labels, such as fluorophores, probes, and dyes, complicate sample preparation and increase cytotoxicity in the process. Therefore, label-free detection becomes essential for in vivo research. The presented 64-channel detection system is designed for label-free detection with the ability to record feeble emissions. Two linear photodetector devices are included in the system, extending the wavelength detection range to 366-680 nm with an improved spectral resolution at an average of 4.9 nm. The performance of the system was validated by detecting unlabeled human hepatocytes (L-02) and other cell-level biologic samples. In addition, the 64-channel detection system was also used for particle counting with a quartz microfluidic chip. The counting method is based on fluorescence spectra differs from those of other devices (i.e., flow cytometry and cell-sorting equipment), which use isolated irradiance intensities.
建立了用于细胞水平激光诱导荧光(LIF)检测的 64 通道检测系统,并将其应用于单事件计数。通常,细胞水平的荧光检测需要染色标签来增强用于光电探测器的散射光强度。然而,染色标签,如荧光染料、探针和染料,会使样品制备复杂化,并在过程中增加细胞毒性。因此,对于体内研究,无标记检测变得至关重要。本研究提出的 64 通道检测系统专为无标记检测而设计,具有记录微弱发射的能力。该系统包括两个线性光电探测器装置,将波长检测范围扩展至 366-680nm,并在平均 4.9nm 的情况下提高了光谱分辨率。通过检测未经标记的人肝细胞(L-02)和其他细胞水平的生物样本,验证了该系统的性能。此外,该 64 通道检测系统还用于带有石英微流控芯片的粒子计数。该计数方法基于与其他设备(即流式细胞仪和细胞分选设备)不同的荧光光谱,这些设备使用孤立的辐照度强度。