Sen Rajannya, Hirvonen Liisa M, Zhdanov Alexander, Svihra Peter, Andersson-Engels Stefan, Nomerotski Andrei, Papkovsky Dmitri
School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland.
These authors contributed equally.
Biomed Opt Express. 2019 Dec 5;11(1):77-88. doi: 10.1364/BOE.11.000077. eCollection 2020 Jan 1.
The properties of a novel ultra-fast optical imager, Tpx3Cam, were investigated for macroscopic wide-field phosphorescent lifetime imaging (PLIM) applications. The camera is based on a novel optical sensor and Timepix3 readout chip with a time resolution of 1.6 ns, recording of photon arrival time and time over threshold for each pixel, and readout rate of 80 megapixels per second. In this study, we coupled the camera to an image intensifier, a 760 nm emission filter and a 50 mm lens, and with a super-bright 627nm LED providing pulsed excitation of a 18 × 18 mm sample area. The resulting macro-imager with compact and rigid optical alignment of its main components was characterised using planar phosphorescent O sensors and a resolution plate mask. Several acquisition and image processing algorithms were evaluated to optimise the system resolution and performance for the wide-field PLIM, followed by imaging a variety of phosphorescent samples. The new PLIM system looks promising, particularly for phosphorescence lifetime-based imaging of O in various chemical and biological samples.
研究了一种新型超快速光学成像仪Tpx3Cam的特性,用于宏观宽场磷光寿命成像(PLIM)应用。该相机基于一种新型光学传感器和Timepix3读出芯片,时间分辨率为1.6 ns,可记录每个像素的光子到达时间和过阈值时间,读出速率为每秒80兆像素。在本研究中,我们将相机与图像增强器、760 nm发射滤光片和50 mm镜头耦合,并使用超亮627 nm LED对18×18 mm的样品区域进行脉冲激发。使用平面磷光O传感器和分辨率板掩模对所得的宏观成像仪进行了表征,其主要组件具有紧凑且刚性的光学对准。评估了几种采集和图像处理算法,以优化宽场PLIM的系统分辨率和性能,随后对各种磷光样品进行成像。新的PLIM系统看起来很有前景,特别是对于各种化学和生物样品中基于磷光寿命的O成像。