Molecular Imaging of Lipid Peroxyl Radicals in Living Cells with a BODIPY−α-Tocopherol Adduct
Bibliographic record
Abstract
An increasing number of reports discuss the role reactive oxygen species (ROS) have in cellular pathologies and cellular signaling processes. Critical to elucidating the underlying chemical mechanism behind these biological processes is the development of novel sensors and reporters with chemical sensitivity and, more importantly, molecular specificity, enabling the spatial and temporal monitoring of a specific ROS concentration in live cells. Here we report for the first time on the application of BODIPY-alpha-Tocopherol adduct (B-TOH), a novel lipophilic fluorescent antioxidant indicator, toward detection of peroxyl radicals in model lipid membranes and their imaging in the lipid membrane of live cells. Studies conducted in model lipid membranes show a 5-fold fluorescence enhancement upon reaction of liposome-embedded B-TOH with peroxyl radicals. The enhancement is independent of the solution pH and membrane composition. In studies in live cells performed under states of growth factor withdrawal and increased oxidative stress, a significant increase in B-TOH emission was also observed. Exogenous sources of free radicals were utilized herein, namely, N,N'-dimethyl-4,4'-bipyridinium dichloride (also known as methyl viologen or paraquat) and uncoated nonemissive CdTe nanoparticles, a source of Cd(2+). The recorded fluorescence intensity of B-TOH was proportional to the concentration of the dye and to the level of cellular oxidative stress. By employing fluorescent dyes such as Lysotracker and Nile Red, we demonstrate the formation of peroxyl radicals in subcellular locations in rat pheochromocytoma (PC12 cells) and in primary mouse hippocampal neural cells under oxidative stress conditions. Specifically, we observed peroxyl radicals in lysosomes. The assessment of the subcellular distribution of B-TOH in living cells deprived from growth factors and/or under oxidative stress may be useful in the future in determining subcellular sites of lipid peroxidation. In summary, results from this study underscore the potential of B-TOH as a sensitive and specific probe enabling the molecular imaging of peroxyl radicals in the lipid membranes of live cells.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".