Ke Ning,Yuanyuan Xie,Wen Sun,Lingke Feng,Can Fang,Rong Pan,Yan Li,Ling Yu
For traditional laboratory microscopy observation, the multi-dimensional, real-time, in situ observation of three-dimensional (3D) tumor spheroids has always been the pain point in cell spheroid observation. In this study, we designed a side-view observation petri dish/device that reflects light, enabling in situ observation of the 3D morphology of cell spheroids using conventional inverted laboratory microscopes. We used a 3D-printed handle and frame to support a first-surface mirror, positioning the device within a cell culture petri dish to image cell spheroid samples. The imaging conditions, such as the distance between the mirror and the 3D spheroids, the light source, and the impact of the culture medium, were systematically studied to validate the in situ side-view observation. The results proved that placing the surface mirror adjacent to the spheroids enables non-destructive in situ real-time tracking of tumor spheroid formation, migration, and fusion dynamics. The correlation between spheroid thickness and dark core appearance under light microscopy and the therapeutic effects of chemotherapy doxorubicin and natural killer cells on spheroids’ 3D structure was investigated.
Yuan Xiang,Zhang Huiling,Cao Huijuan,Mao Guojiang,Wei Lin
A visible and sensitive assay for the quantitative detection of β-glucosidase (β-glu) activity based on Au@CeO 2 core-shell nanoparticles (Au@CeO 2 NPs) is described. As a hydrolytic enzyme, β-glu can promote the hydrolysis of β-arbutin to hydroquinone (HQ), which can trigger the decomposition of the CeO 2 shell. With the single-particle enumeration (SPE) strategy coupled with dark field optical microscopy (DFM), an obvious color alteration of single Au@CeO 2 NPs during the etching process can be observed in real-time. By statistically calculating the number of the etched nanoparticles, the β-glu activity level can be quantified accurately. This assay displays a broad linear range from 0.5 to 50 mU⋅mL −1 and low detection limit of 0.12 mU⋅mL −1 . In addition, this method was successfully used to determine β-glu in real samples and acquires satisfactory recoveries in the range of 97.1-102.0%. This study provides a visualization analysis method for β-glu, which may be helpful for monitoring other targets in the future. Graphical Abstract
危险品化学品经营许可证（带存储）