Mitochondria-Assisted Photooxidation to Track Singlet Oxygen at Homeostatic Membrane Microviscosity

Abstract

Using intracellular-controlled photochemistry to track dynamic organelle processes is gaining attention due to its broad applications. However, most of the employed molecular probes usually require toxic photosensitizers and complex bioanalytical protocols. Here, the synthesis and performance of two new subcellular probes (MitoT1 and MitoT2) are described. The probes undergo photooxidation in the damaged tissue of zebrafish, a model system for tissue regeneration studies. Using high-resolution confocal microscopy and fluorescence spectroscopy, we combine the mentioned photoinduced interconversion at the homeostatic membrane viscosity to track singlet oxygen activity selectively. The continuous and real-time biosensing method reported here provides a new approach for simultaneously detecting endogenous singlet oxygen and viscosity status.

Unveiling cellular vitality: peptide fluorescent probes illuminate mitochondrial dynamics and ROS activity

“…2. According to previous literature reports, 13,17–19 and the corresponding mass spectrometry (HRMS) studies, this is assigned to the formation of dioxetane and aldehyde species promoted by singlet oxygen photooxygenation (see Fig. S5, ESI† file), thus activating the green-emission response channel.…”

“…5. Zebrafish serves as a model system for studying tissue regeneration, 19 aiding in the comprehension of tissue maintenance and turnover processes. The wounded area is predominantly characterized by dead cells.…”

Fluorescent Probe for in Vivo Partitioning into Dynamic Lipid Droplets Enables Monitoring of Water Permeability-Induced Edema

“…32 The challenge in edema monitoring implies several requirements, fundamentally: (1) to stain not only internal yolk lipids but epidermal yolk cells (platelets) as well, (2) to respond to an osmotic imbalance, (3) to allow continuous monitoring (stable signal response overtime) and, (4) to have an internal reference signal or second response channel. 33 Fluorescent probes able to overcome these features using single-molecule assays have not been reported to date, and commercial fluorescent reporters only fulfill points 3 and 4. Bioanalytical protocols using transgenic zebrafish models expressing fluorescent proteins for LDs 9 together with yolk platelet markers could be a viable dual strategy.…”

Exploring mitochondrial targeting: an innovative fluorescent probe reveals Nernstian potential and partitioning combination

“…Therefore, key optimization features for mitochondrial targeting illustrated in Fig. 1 involved functional optimization by ortho-/para-substitution to the benz[e]indolinium to tune the cationic-hydrophobic balance to stain mitochondria, 16,17 thus modifying its dielectric features, resulting in the development of aggregation properties attributed to the interaction of transition dipoles. [18][19][20] The optical optimization through umbelliferone incorporation was used to enhance the fluorescence response.…”