Light-sheet fluorescence microscopy (LSFM) has been widely used to image the three-dimensional (3D) structures and functions of various millimeter-size bio-specimen such as zebrafish. However, the sample adsorption and scattering cause shading of the light-sheet illumination, preventing the even 3D image of thick samples. Herein, we report a continuous-rotational light-sheet microscope (CR-LSM) that enables simultaneous 3D bright-field and fluorescence imaging. With a high-accuracy rotational stage, CR-LSM records the outline projections and the fluorescent images of the sample at multiple rotation angles. Then, 3D morphology and fluorescent structure were reconstructed with a developed algorithm. Using CR-LSM, zebrafish’s whole-fish contour and blood vessel structures were obtained simultaneously.

Three-dimensional (3D) cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models. The modern optical sectioning microscopic methods make it possible to probe the complexity of 3D cell cultures but are limited by the inherent opaqueness. While tissue optical clearing methods have emerged as powerful tools for investigating whole-mount tissues in 3D, they often have limitations, such as being too harsh for fragile 3D cell cultures, requiring complex handling protocols, or inducing tissue deformation with shrinkage or expansion. To address this issue, we proposed a modified optical clearing method for 3D cell cultures, called MACS-W, which is simple, highly efficient, and morphology-preserving. In our evaluation of MACS-W, we found that it exhibits excellent clearing capability in just 10min, with minimal deformation, and helps drug evaluation on tumor spheroids. In summary, MACS-W is a fast, minimally-deformative and fluorescence compatible clearing method that has the potential to be widely used in the studies of 3D cell cultures.

光片显微镜由于具有强大的光学层切能力、较快的成像速度和较低的光损伤，成为三维成像的重要工具。光片显微镜通常利用两个垂直放置的物镜分别进行照明和成像，这带来了对样品的空间限制并禁用了高数值孔径的成像物镜。以倾斜平面照明和微镜微器件反射技术为代表的单物镜光片显微技术突破上述限制，展示出在高分辨率和体积高速成像方面的优势，并且可与超分辨显微术等多种技术结合，在近年来取得了巨大发展。介绍单物镜光片显微成像技术的原理、关键性能的提升和其在生物医学的应用。

Photodynamic therapy (PDT) has limited effects in treating metastatic breast cancer. Immune checkpoints can deplete the function of immune cells; however, the expression of immune checkpoints after PDT is unclear. This study investigates whether the limited efficacy of PDT is due to upregulated immune checkpoints and tries to combine the PDT and immune checkpoint inhibitor to observe the efficacy. A metastatic breast cancer model was treated by PDT mediated by hematoporphyrin derivatives (HpD-PDT). The anti-tumor effect of HpD-PDT was observed, as well as CD4+T, CD8+T and calreticulin (CRT) by immunohistochemistry and immunofluorescence. Immune checkpoints on T cells were analyzed by flow cytometry after HpD-PDT. When combining PDT with immune checkpoint inhibitors, the antitumor effect and immune effect were assessed. For HpD-PDT at 100mW/cm² and 40, 60 and 80J/cm², primary tumors were suppressed and CD4+T, CD8+T and CRT were elevated; however, distant tumors couldn’t be inhibited and survival could not be prolonged. Immune checkpoints on T cells, especially PD1 and LAG-3 after HpD-PDT, were upregulated, which may explain the reason for the limited HpD-PDT effect. After PDT combined with anti-PD1 antibody, but not with anti-LAG-3 antibody, both the primary and distant tumors were significantly inhibited and the survival time was prolonged, additionally, CD4+T, CD8+T, IFN-γ+CD4+T and TNF-α+CD4+T cells were significantly increased compared with HpD-PDT. HpD-PDT could not combat metastatic breast cancer. PD1 and LAG-3 were upregulated after HpD-PDT. Anti-PD1 antibody, but not anti-LAG-3 antibody, could augment the antitumor effect of HpD-PDT for treating metastatic breast cancer.

人类表皮生长因子受体-2（HER2）的异常扩增会导致癌细胞的过度增殖和肿瘤恶化。在采用常规光学显微成像技术检测扩增水平较高的乳腺癌细胞HER2基因时，荧光原位杂交探针的荧光信号斑点呈簇状分布，难以精确计数。应用结构光照明超分辨成像技术对HER2基因荧光原位杂交的病理切片进行成像，从而分辨距离较近的荧光探针。通过大视场扫描成像和图像拼接，对数百个细胞进行成像和统计分析，提高了高扩增水平病理切片上HER2探针计数的准确性。

电动化、智能化的光学显微成像系统需要能够实时测量系统的焦面漂移并进行校正，从而实现对活细胞的长时间观测和全片的病理切片扫描。设计一种探测非对称光束界面反射后的光斑位置从而进行焦面漂移测量的方法。利用ZEMAX软件光学仿真了反射光斑在不同离焦情况下的光斑形状，搭建了集成化的漂移测量模块并在商用正置显微镜上对其进行了验证。结果表明，针对60×浸油物镜，所提系统的漂移测量精度达250 nm，漂移校正的响应时间小于500 ms，满足了高分辨率长时间成像的要求。

基于磷脂酰肌醇 3-激酶（ PI3K）/丝苏氨酸蛋白激酶（ AKT）信号通路探究安罗替尼对 A549细胞增殖、凋亡的影响。将 A549细胞分为对照组、各剂量试验组、安罗替尼组、阳性药物组、抑制剂组和激活剂组。干预 24 h后检测 A549细胞活力、细胞形态、增殖率、凋亡情况及相关蛋白的表达水平。结果显示： 20 μmol/L安罗替尼处理后， A549细胞活力降低；安罗替尼组和阳性药物组较对照组 A549细胞生长受到抑制，细胞增殖率、细胞周期素 D1（Cyclin D1）及 p-PI3K、p-AKT的表达水平降低，细胞凋亡数量和半胱氨酸天冬氨酸蛋白酶 -3（Caspase-3）的表达水平升高。抑制剂增强了上述各指标的变化，而激活剂则具有与抑制剂相反的趋势。该研究结果说明，安罗替尼可显著抑制人肺癌 A549细胞的增殖并促进其凋亡，其作用机制可能与抑制 PI3K/AKT通路的信号转导相关。该研究结果进一步说明了安罗替尼作为抗肺癌药物的潜在价值，为抗肺癌药物的研发提供了新的理论基础。

随着生活和消费水平提高, 消费者对于乳制品食品安全及品质的要求越来越高。 原料奶质量直接影响乳制品的生产与消费安全, 在牛奶收储及生产环节都需要对各种非法添加物进行严格检测, 以保证产品质量。 目前常用的检测方法主要以化学法与仪器分析方法为主, 需要针对不同类型添加物设计前处理步骤, 过程繁琐, 检测效率低, 无法满足实时在线需要。 针对多种类掺假异常牛奶样品实时在线检测需要, 研究了基于中红外光谱的非靶向检测方法。 实验样品选择蒙牛公司六个奶质稳定的奶源地收集到的天然原奶样品, 并配制含有多种掺假物的异常牛奶样品集。 采集样品中红外光谱, 并针对在线检测过程中的干扰来源, 选择平滑滤波、 多元散射校正、 基线校正及归一化等预处理方法, 提高光谱信噪比与一致性。 为了提高非靶向模型识别准确度及稳健性, 根据牛奶样品中红外光谱特征, 选择无信息变量消除（MC-UVE）、 无变量信息消除-连续投影（UVE-SPA）与竞争自适应重加权采样（CARS）三种方法, 筛选原始光谱中的特征波长变量。 在得到的不同特征波长变量组合的基础上, 分别建立基于偏最小二乘判别（PLS-DA）及支持向量机（SVM）的鉴别模型, 对多种掺假物异常牛奶样品进行非靶向鉴别。 实验结果表明, SVM模型鉴别准确度优于PLS-DA, CARS方法筛选得到的变量组合应用于不同鉴别模型的效果均较优, 与SVM模型结合对训练集与测试集的分类准确率分别达到97.84%与94.55%。 分析特征波长变量分布可知, CARS方法筛选出的变量主要集中在异常牛奶样品光谱特征比较明显的区域。 样品误分类结果表明, 该模型组合可以较为准确识别异常牛奶样品, 具有较好的特异性。 研究结果表明, 基于红外光谱技术建立非靶向鉴别模型可以实现多种异常牛奶样品快速准确识别, 为牛奶掺假及生产过程在线检测提供了支持。