超分辨荧光显微镜突破了光学衍射极限造成的空间分辨率限制，使得生物学家能够在生命体和细胞具有活性的状态下，对其功能与结构进行高精度动态记录，有望揭示更多重要的生命现象细节。然而，由于超分辨荧光显微技术的成像视场、深度、分辨率、速度等不易兼得，所以解卷积作为一种最有效且直接的求解逆问题的框架，被广泛应用于增强超分辨显微镜的时空分辨率。研究人员聚焦于通过相应算法设计实现高质量显微图像的重建，在一定程度上克服了超分辨荧光显微镜的硬件限制，可以更好地恢复生物信息。本文首先介绍了解卷积方法的基本原理及其发展历程，接着列举了不同解卷积技术在不同模态下的重建原理和效果以及这些技术在生物学上的应用，最后总结了基于深度学习的解卷积方法在超分辨荧光显微镜技术上的最新进展和未来的发展潜力，并对包括傅里叶环相关的定量评估图像重建质量的方法的最新进展进行了阐述。

Single-cell volumetric imaging is essential for researching individual characteristics of cells. As a nonscanning imaging technique, light field microscopy (LFM) is a critical tool to achieve real-time three-dimensional imaging with the advantage of single-shot. To address the inherent limits including nonuniform resolution and block-wise artifacts, various modified LFM strategies have been developed to provide new insights into the structural and functional information of cells. This review will introduce the principle and development of LFM, discuss the improved approaches based on hardware designs and 3D reconstruction algorithms, and present the applications in single-cell imaging.Single-cell volumetric imaging is essential for researching individual characteristics of cells. As a nonscanning imaging technique, light field microscopy (LFM) is a critical tool to achieve real-time three-dimensional imaging with the advantage of single-shot. To address the inherent limits including nonuniform resolution and block-wise artifacts, various modified LFM strategies have been developed to provide new insights into the structural and functional information of cells. This review will introduce the principle and development of LFM, discuss the improved approaches based on hardware designs and 3D reconstruction algorithms, and present the applications in single-cell imaging.

Deviation of the H+ concentration from optimum values within the organelles is closely associated with irregular cellular functions that cause the onset of various diseases. Therefore, determining subcellular pH values in live cells and tissues is valuable for diagnostic purposes. In this study, we report a novel ratiometric fluorescence probe 1H-pyrazole-3-carboxylic acid, 4-(benzo[d]thiazol-2-yl)-3-(2,4-dihydroxy-3-methylphenyl)-1H-pyrazole-5-carboxylicacid4-(2-benz othiazolyl)-5-(2,4-dihydroxy-3-methylphenyl), to which we will refer as ThiAKS Green (Thiazole AKyol shifting green), that is pH sensitive. The results presented here show that the probe can penetrate the cell membrane in less than 30 minutes and does not show any detectable toxicity. The measured color shifts up on pH change are linear and most significant around physiological pH (pKa=7.45), thus making this probe suitable for live-cell imaging and intracellular pH measurements. During the long-incubation periods following the application of the probe and the fluorescent microscopy measurements, it shows stable properties and is easy to detect in live cells. In conclusion, the results suggest that ThiAKS Green can be used to obtain precise information on the H+ distribution at various compartments of the live cells.

探索合成方法简单的近红外比率荧光探针具有重要意义。本文以高山榕树叶为碳源，乙醇、丙酮为提取剂，采用溶剂热法于200 ℃高压反应釜中反应10 h，一步合成具有近红外双发射荧光特性的碳量子点(CDs)。透射电镜(TEM)表明合成的CDs分散性好、平均尺寸约为4.70 nm，晶格常数为0.32 nm。X射线衍射(XRD)结果显示该碳点在2θ=22°附近有一个较宽的衍射峰，表明该CDs为sp2杂化的无定形碳。傅里叶红外光谱(FT-IR)显示其含有氨基、羰基、醇羟基和甲基。荧光光谱表明，最大激发波长410 nm，最大发射波长为466 nm和676 nm。以硫酸奎宁做参比测得荧光量子产率为26.31%。实验还探讨了CDs的酸碱、光学、抗盐和金属离子的稳定性以及细胞成像效果。以上结果表明，该碳点具有原料廉价易得、制作简便、对酸碱稳定、抗光漂白性良好等优点，具有良好的光学成像应用前景。

为了同时满足较大的视场和较高分辨率的需求, 开发了一套全息无透镜显微成像系统和配套算法, 实现对微米级样品的无透镜显微成像。搭建了一套由LED光源、针孔、被测样品与CMOS图像传感器组成的全息无透镜显微成像系统, 并对针孔直径、成像面尺寸、光源到样品的距离, 以及样品面到CMOS图像传感器的距离进行了优化。其次, 开发了从系统采集的全息图中恢复样品图像的角谱法算法。最后, 使用该成像系统和配套算法, 分别对具有微米级结构分辨率测试靶, 和肺癌细胞悬浮液进行了显微成像。该全息无透镜显微成像系统的分辨率为4.4 μm, 成像视场尺寸为5.7 mm×4.3 mm, 实现了微米级结构和肺癌细胞较清晰的显微成像。全息无透镜显微成像系统结构简单、无像差干扰, 可以实现大视场下较高分辨率的显微成像。

捕捉细胞内分子活动发生的动态过程, 从细胞分裂到囊泡运输再到胞内钙离子浓度变化等大量快速发生和发展的生理过程是很多科研工作者的需要。这个过程不仅要求较高的时间分辨率, 还要求较低的激发光强度以使样品的淬灭和光损伤达到最小。因此, 相机的高灵敏度图像传感器起到了决定作用。鉴于近几年图像传感器发展迅速, 本文对其进行了系统的阐述, 综述了相机的图像传感器技术上的最新突破和改进。进而, 总结了其在活细胞成像中的应用, 对比了两种主流传感器并展望了未来发展趋势。