研究目的
To develop a novel fluorescent probe for specific and real-time monitoring of lysosomal processes, addressing limitations of existing probes such as low specificity, high cytotoxicity, and poor photostability.
研究成果
2M-DPAS is a highly effective fluorescent probe with AIE characteristics, offering good biocompatibility, high selectivity, bright emission, pH-independence, and excellent photostability. It enables real-time monitoring of lysosomal dynamics, including autophagy, mitophagy, and phagocytosis of macromolecules, making it a promising tool for studying lysosome-related diseases like Alzheimer's.
研究不足
The study is limited to in vitro cell cultures (HeLa and HMCs); in vivo applications and broader cell types were not explored. The probe's performance in complex biological environments and long-term effects require further investigation.
1：Experimental Design and Method Selection:
The study involved designing and synthesizing a novel AIE fluorogen, 2M-DPAS, based on DPAS, to target lysosomes due to its morpholine groups and AIE properties. Methods included synthesis, photophysical property studies, cell staining, co-staining with commercial dyes, photostability tests, cytotoxicity assays, and monitoring of autophagy, mitophagy, and phagocytosis processes.
2：Sample Selection and Data Sources:
HeLa cells and human meningeal cells (HMCs) were used as cell models. Samples included 2M-DPAS, Lysotracker Red (LTR), MitoTracker Red (MTR), FAM-Aβ, rapamycin, and 3-MA.
3：List of Experimental Equipment and Materials:
Confocal microscopy for imaging, MTT assay kit for cytotoxicity, various chemicals and dyes as specified.
4：Experimental Procedures and Operational Workflow:
Cells were stained with probes, treated with inhibitors or inducers, and imaged over time. Procedures included incubation, washing, and fluorescence measurement under controlled conditions.
5：Data Analysis Methods:
Fluorescence intensity was quantified, Pearson's correlation coefficient was calculated for specificity, and statistical analysis was performed for cytotoxicity and photostability.
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