Tracking mitochondrial Cu(I) fluctuations through a ratiometric fluorescent probe in AD model cells: Towards understanding how AßOs induce mitochondrial Cu(I) dyshomeostasis.

Mitochondrial copper signaling pathway plays a role in Alzheimer's disease (AD), especially in relevant Amyloid-ß oligomers (AßOs) neurotoxicity and mitochondrial dysfunction. Clarifying the relationship between mitochondrial copper homeostasis and both of mitochondrial dysfunction and AßOs neurotoxicity is important for understanding AD pathogenesis. Herein, we designed and synthesized a ratiometric fluorescent probe CHC-NS4 for Cu(I). CHC-NS4 possesses excellent ratiometric response, high selectivity to Cu(I) and specific ability to target mitochondria. Under mitochondrial dysfunction induced by oligomycin, mitochondrial Cu(I) levels gradually increased, which may be related to inhibition of ATP7A-mediated Cu(I) exportation and/or high expression of COX. On this basis, CHC-NS4 was further utilized to visualize the fluctuations of mitochondrial Cu(I) levels during progression of AD model cells induced by AßOs. It was found that mitochondrial Cu(I) levels were gradually elevated during the AD progression, which depended on not only AßOs concentration but also incubation time. Moreover, endocytosis maybe served as a prime pathway mode for mitochondrial Cu(I) dyshomeostasis induced by AßOs during AD progression. These results have provided a novel inspiration into mitochondrial copper biology in AD pathogenesis.

Single nanowire-based fluorescence lifetime thermometer for simultaneous measurement of intra- and extra-cellular temperatures.

A silicon nanowire-based fluorescence lifetime thermometer (NWFLT) was fabricated for the simultaneous measurement of intra- and extra-cellular temperatures. Using the NWFLT, an obvious heterogeneity of the temperature was found along the longitude direction of the NWFLT, especially between the inside and outside of the cell.

Simultaneous Monitoring of the Adenosine Triphosphate Levels in the Cytoplasm and Nucleus of a Single Cell with a Single Nanowire-Based Fluorescent Biosensor.

Simultaneous monitoring of the ATP levels at various sites of a single cell is crucial for revealing the ATP-related processes and diseases. In this work, we rationally fabricated single nanowire-based fluorescence biosensors, by which the ATP levels of the cytoplasm and nucleus in a single cell can be simultaneously monitored with a high spatial resolution. Utilizing the as-fabricated single nanowire biosensor, we demonstrated that the ATP levels of the cytoplasm were around 20-30% lower than that of the nucleus in both L929 and HeLa cells. Observing the ATP fluctuation of the cytoplasm and nucleus of the L929 and HeLa cells stimulated by Ca2+, oligomycin, or under cisplatin-induced apoptosis, we found that the ATP levels at two cellular sites exhibited discriminative changes, revealing the different mechanisms of the ATP at these two cellular sites in response to the stimulations.

Sensitive and Stable Thermometer Based on the Long Fluorescence Lifetime of Au Nanoclusters for Mitochondria.

Detecting the temperature of intracellular mitochondria with high sensitivity and stability is crucial to understanding the cellular metabolism and revealing the processes of mitochondria-related physiology. In this paper, employing the long fluorescence lifetime of modified Au nanoclusters (mAuNCs) by 4-(carboxybutyl) triphenylphosphonium bromide, we developed a fluorescence lifetime thermometer with high sensitivity and stability for the temperature of the intracellular mitochondria. A high relative temperature sensitivity of 2.8% and excellent photostability were achieved from the present thermometer. After incubation with L929 cells, the mAuNCs could be endocytosed into the cells and targeted the mitochondria, and the temperature changes at the L929 cells' mitochondria, which were stimulated by carbonyl cyanide 3-chlorophenylhydrazone and Ca2+, were successfully detected via the fluorescence lifetime images of the mAuNCs. Furthermore, utilizing the mAuNCs, we clarified the effect of Mg2+ on the temperature of the intracellular mitochondria. The strategy of employing a material with a long fluorescence lifetime and remarkable stability to fabricate the fluorescence lifetime thermometer for mitochondria can be used to design various thermometers for other organelles.