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用于时间分辨荧光光谱学的256×2 SPAD线传感器。

256 × 2 SPAD line sensor for time resolved fluorescence spectroscopy.

作者信息

Krstajić Nikola, Levitt James, Poland Simon, Ameer-Beg Simon, Henderson Robert

出版信息

Opt Express. 2015 Mar 9;23(5):5653-69. doi: 10.1364/OE.23.005653.

Abstract

We present a CMOS chip 256 × 2 single photon avalanche diode (SPAD) line sensor, 23.78 µm pitch, 43.7% fill factor, custom designed for time resolved emission spectroscopy (TRES). Integrating time-to-digital converters (TDCs) implement on-chip mono-exponential fluorescence lifetime pre-calculation allowing timing of 65k photons/pixel at 200 Hz line rate at 40 ps resolution using centre-of-mass method (CMM). Per pixel time-correlated single-photon counting (TCSPC) histograms can also be generated with 320 ps bin resolution. We characterize performance in terms of dark count rate, instrument response function and lifetime uniformity for a set of fluorophores with lifetimes ranging from 4 ns to 6 ns. Lastly, we present fluorescence lifetime spectra of multicolor microspheres and skin autofluorescence acquired using a custom built spectrometer. In TCSPC mode, time-resolved spectra are acquired within 5 minutes whilst in CMM mode spectral lifetime signatures are acquired within 2 ms for fluorophore in cuvette and 200 ms for skin autofluorescence. We demonstrate CMOS line sensors to be a versatile tool for time-resolved fluorescence spectroscopy by providing parallelized and flexible spectral detection of fluorescence decay.

摘要

我们展示了一款定制设计用于时间分辨发射光谱(TRES)的CMOS芯片,它是256×2单光子雪崩二极管(SPAD)线传感器,像素间距为23.78 µm,填充因子为43.7%。集成的时间数字转换器(TDC)实现了片上单指数荧光寿命预计算,使用质心法(CMM)在200 Hz行速率下以40 ps分辨率对每个像素65k个光子进行计时。每个像素的时间相关单光子计数(TCSPC)直方图也可以以320 ps的bin分辨率生成。我们针对一组寿命范围为4 ns至6 ns的荧光团,对暗计数率、仪器响应函数和寿命均匀性等性能进行了表征。最后,我们展示了使用定制构建的光谱仪采集的多色微球荧光寿命光谱和皮肤自发荧光光谱。在TCSPC模式下,对于比色皿中的荧光团,5分钟内可采集时间分辨光谱,而对于皮肤自发荧光,在CMM模式下200 ms内可采集光谱寿命特征。我们通过提供并行且灵活的荧光衰减光谱检测,证明了CMOS线传感器是时间分辨荧光光谱的一种通用工具。

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