Computational Photon Counting Using Multithreshold Peak Detection for Fast Fluorescence Lifetime Imaging Microscopy.
Computational Photon Counting Using Multithreshold Peak Detection for Fast Fluorescence Lifetime Imaging Microscopy.
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DOI：
10.1021/acsphotonics.2c00505
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发表时间：
2022-08-17
期刊：
影响因子：
7
通讯作者：
Boppart, Stephen A.
中科院分区：
文献类型：
--
作者：
Sorrells, Janet E.;Iyer, Rishyashring R.;Yang, Lingxiao;Martin, Elisabeth M.;Wang, Geng;Tu, Haohua;Marjanovic, Marina;Boppart, Stephen A.
时间分辨光子计数方法具有有限的带宽，这限制了荧光寿命成像显微镜（FLIM）等技术的采集速度。为了实现更快的成像，当检测器的输出以高采样率直接数字化时，可以采用计算方法来计数光子。在这里，我们提出了计算光子计数使用混合光电探测器结合多阈值峰值检测计数的情况下，一个或多个光子到达检测器内的检测器响应时间。这种方法可以用来区分多达五个光子计数每个数字化点，而以前的演示计算光子计数的数据与光电倍增管只计数一个光子的时间。我们在两个自由移动的C。elegans和人乳腺癌细胞系进行凋亡，这种新的多阈值峰值检测方法可以准确地表征强度和荧光寿命的样品产生的光子率高达223%，高于以前证明的光子计数FLIM系统。
Time-resolved photon counting methods have a finite bandwidth that restricts the acquisition speed of techniques like fluorescence lifetime imaging microscopy (FLIM). To enable faster imaging, computational methods can be employed to count photons when the output of a detector is directly digitized at a high sampling rate. Here, we present computational photon counting using a hybrid photodetector in conjunction with multithreshold peak detection to count instances where one or more photons arrive at the detector within the detector response time. This method can be used to distinguish up to five photon counts per digitized point, whereas previous demonstrations of computational photon counting on data acquired with photomultiplier tubes have only counted one photon at a time. We demonstrate in both freely moving C. elegans and a human breast cancer cell line undergoing apoptosis that this novel multithreshold peak detection method can accurately characterize the intensity and fluorescence lifetime of samples producing photon rates up to 223%, higher than previously demonstrated photon counting FLIM systems.
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