A wide-field microscope with epi-fluorescence and selective plane illumination was combined with a single-photon avalanche diode (SPAD) array camera to enable live-cell fluorescence lifetime imaging (FLIM) using time-correlated single-photon counting (TCSPC). The camera sensor comprised of \(\text {192}\times \text {128}\) pixels, each integrating a single SPAD and a time-to-digital converter. Jointly, they produced a stream of single-photon images of photon arrival times with \(\approx \text {38 ps}\) accuracy. The photon arrival times were subject to systematic delays and nonlinearities, which were corrected by a Monte-Carlo algorithm. The SPAD camera was then applied to FLIM where histogramming the resulting photon arrival times in each pixel resulted in decays compatible with common data processing pipelines for fluorescence lifetime analysis. The capabilities of the TCSPC camera-based FLIM microscope were demonstrated by imaging living unicellular photosynthetic algae and artificial lipid vesicles. Epi-fluorescence illumination enabled rapid fluorescence lifetime imaging of living cells and selective-plane illumination enabled 3-dimensional FLIM of stationary samples.

具有落射荧光和选择性平面照明的宽视场显微镜与单光子雪崩二极管 (SPAD) 阵列相机相结合，可使用时间相关单光子计数 (TCSPC) 进行活细胞荧光寿命成像 (FLIM)。相机传感器由\(\text {192}\times \text {128}\)像素组成，每个像素集成一个 SPAD 和一个时间数字转换器。他们共同生成了光子到达时间的单光子图像流，精度为\(\approx \text {38 ps}\)。光子到达时间受到系统延迟和非线性的影响，并通过蒙特卡罗算法进行校正。然后将 SPAD 相机应用于 FLIM，其中对每个像素中产生的光子到达时间进行直方图绘制，从而产生与荧光寿命分析的常用数据处理流程兼容的衰减。基于 TCSPC 相机的 FLIM 显微镜的功能通过对活体单细胞光合藻类和人工脂质囊泡进行成像来证明。落射荧光照明可实现活细胞的快速荧光寿命成像，选择性平面照明可实现固定样品的 3 维 FLIM。