Multiplexed imaging detection of live cell intracellular changes in early apoptosis with aggregation-induced emission fluorogens
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Apoptosis is an important process for maintaining tissue homeostasis and eliminating abnormal cells in multicellular organisms. Abnormality in apoptosis often leads to severe diseases such as cancers. Better understanding of its mechanisms and processes is therefore important. Accompanying molecular biology events of apoptosis is a series of cellular morphology changes: nucleus condensation, cell shrinkage and rounding, cell surface blebbing, dynamic blebbing, apoptotic membrane protrusions and nucleus fragmentations and finally, the formation and release of apoptotic bodies. It is difficult to detect cellular changes in the early phase of apoptosis due to the subtle changes at this phase. In the current study, we induced apoptosis in HeLa cells with H2O2 and used nuclear dye Hoechst 33258, mitochondria, lysosome and cytoplasmic protein specific aggregation-induced emission fluorogens (AIEgens), TPE-Ph-In, 2M-DABS and BSPOTPE to successfully perform live cell multiplexed imaging to investigate early apoptosis cellular events. We showed the gradual dissipation of mitochondria membrane potential until it is nondetectable by TPE-Ph-In. Increased mitophagy detected by TPE-Ph-In and 2M-DABS, condensed nucleus detected by Hoechst 33258, increased permeability and/or reduced integrity of nuclear membrane, and increased intracellular vesicles detected by 2M-DABS are some of the early events of apoptosis.
Keywordsapoptosis multiplexed imaging HeLa aggregation-induced emission fluorogens (AIEgens)
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This work was supported by the Hong Kong Branch of Chinese National Engineering Research Centres for Tissue Restoration and Reconstruction. We acknowledged the use of South Australian nodes of the Australian Microscopy & Microanalysis Research Facility and the Australian National Fabrication Facility at Flinders University.
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