A mitochondria-specific fluorescent probe for rapidly assessing cell viability
Jianmei Chen1, Yurui Xu1, Ya Gao1
1National Laboratory of Solid State Microstructures, Chemistry and Biomedicine Innovation Center, College of Engineering and Applied Sciences, Jiangsu Key Laboratory of Artificial Functional Materials, Nanjing University, Nanjing, 210093, P. R. China.
Abstract:
Rapid detection of cell viability is of importance for various aspects of basic and applied research. However, the most commonly used MTT assay suffers from some disadvantages of time-consuming, poor solubility of formazan crystals, and interference of cell extracts, resulting in either delayed results or inconsistent cell viability. Accumulating evidences show that mitochondrial activity is closely associated with cell proliferation and apoptosis, making it valuable for real-time monitoring of cell fates. Herein, we present a novel type of cell-permeant mitochondrial dyes, composed of coumarin-quinoline conjugates (CQCs), for assessing cell viability through monitoring mitochondrial dynamic changes. CQCs exhibited superior preference for mitochondria, and could sensitively monitor their fissions under pathological conditions. Particularly, CQCs could accurately assess the cell viability within 10 min, allowing for rapidly evaluating therapeutic effects with high sensitivity and accuracy, in comparison of MTT assay. Therefore, CQCs provide a proof-of-concept assay that is appropriately practical either independently or in complementation with other assays for the analysis of cellular viability in broad range of biomedical applications.
Insights
Rapidly assess cell viability using novel coumarin-quinoline conjugates (CQCs). These mitochondrial dyes offer a fast, sensitive alternative to the MTT assay for real-time monitoring of cell health in biomedical research.
Area of Science:
- Biochemistry
- Cell Biology
- Biomedical Engineering
Background:
- Cell viability assays are crucial for research but common methods like the MTT assay are slow and unreliable.
- Mitochondrial activity correlates with cell proliferation and apoptosis, offering a target for real-time monitoring.
- Existing assays face challenges including time consumption, poor formazan solubility, and interference from cell extracts.
Purpose of the Study:
- To develop a novel, rapid, and sensitive method for assessing cell viability.
- To utilize coumarin-quinoline conjugates (CQCs) as cell-permeant mitochondrial dyes for monitoring mitochondrial dynamics.
- To establish CQCs as a practical tool for evaluating therapeutic effects and cellular health.
Main Methods:
- Synthesis of coumarin-quinoline conjugates (CQCs) as novel mitochondrial dyes.
- Assessment of CQC mitochondrial targeting and sensitivity to mitochondrial fission.
- Comparison of CQC-based viability assessment with the MTT assay for speed and accuracy.
Main Results:
- CQCs demonstrated high specificity and preference for mitochondria.
- CQCs effectively monitored mitochondrial dynamics, including fissions under pathological conditions.
- Cell viability was accurately assessed by CQCs within 10 minutes, significantly faster than the MTT assay.
- CQCs showed high sensitivity and accuracy in evaluating therapeutic effects.
Conclusions:
- Coumarin-quinoline conjugates (CQCs) offer a rapid and sensitive method for cell viability assessment.
- CQCs provide a valuable tool for real-time monitoring of cell fates by tracking mitochondrial dynamics.
- This novel assay is practical for biomedical applications, either alone or in combination with other methods.
