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<p>Abnormal alterations in mitochondrial viscosity, an important biophysical indicator of the mitochondrial microenvironment and functional health, are strongly linked to the initiation and progression of many diseases. Herein, a series of fluorescent probes (MPS1–MPS5) targeting mitochondria were designed and synthesized featuring adjustable alkyl chain lengths, which exhibit high sensitivity to viscosity and enable precise monitoring of mitochondrial dysfunction. We systematically examined how varying the chain length affects the photophysical properties of these probes. Our results revealed that increasing the length of the alkyl chain on the pyridine nitrogen enhanced the probes’ sensitivity to viscosity. Among the series, MPS5, which incorporates a pentyl group and operates based on the twisted intramolecular charge transfer (TICT) mechanism, exhibited the most significant fluorescence enhancement, making it the most effective probe for detecting changes in mitochondrial viscosity. Notably, MPS5 exhibited excellent anti-interference capability, enabling precise mitochondrial imaging without the need for washing. Experimental data confirmed that MPS5 can distinctly differentiate mitochondrial viscosity under various physiological states, offering a powerful tool for studying cellular energy metabolism and dysfunction. Moreover, MPS5 was found to be capable of ultra-low concentration imaging, achieving a detection limit as low as 50 nM. Subsequent studies also demonstrated that MPS5 could effectively assess the impact of different drug interventions on mitochondrial viscosity abnormalities in disease models, highlighting its potential for drug efficacy evaluation. Notably, MPS5 also enables the assessment of acute alcoholic liver injury in mouse models at the organ level.</p>

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Design of wash-free fluorescent probes with alkyl chain length modulation strategy for mitochondrial viscosity detection and alcoholic liver injury diagnosis

  • 期刊论文

  • 发表日期：

    2026年5月23日

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  • Sichen Zhang,
  • Lifeiyang Wan,
  • Cheng Huang,
  • Yunjia Zhao,
  • Xinyue Zhao,
  • Kaiyuan Liu,
  • Hui Wang,
  • Ping Li,
  • Lei Hu

摘要

Abnormal alterations in mitochondrial viscosity, an important biophysical indicator of the mitochondrial microenvironment and functional health, are strongly linked to the initiation and progression of many diseases. Herein, a series of fluorescent probes (MPS1–MPS5) targeting mitochondria were designed and synthesized featuring adjustable alkyl chain lengths, which exhibit high sensitivity to viscosity and enable precise monitoring of mitochondrial dysfunction. We systematically examined how varying the chain length affects the photophysical properties of these probes. Our results revealed that increasing the length of the alkyl chain on the pyridine nitrogen enhanced the probes’ sensitivity to viscosity. Among the series, MPS5, which incorporates a pentyl group and operates based on the twisted intramolecular charge transfer (TICT) mechanism, exhibited the most significant fluorescence enhancement, making it the most effective probe for detecting changes in mitochondrial viscosity. Notably, MPS5 exhibited excellent anti-interference capability, enabling precise mitochondrial imaging without the need for washing. Experimental data confirmed that MPS5 can distinctly differentiate mitochondrial viscosity under various physiological states, offering a powerful tool for studying cellular energy metabolism and dysfunction. Moreover, MPS5 was found to be capable of ultra-low concentration imaging, achieving a detection limit as low as 50 nM. Subsequent studies also demonstrated that MPS5 could effectively assess the impact of different drug interventions on mitochondrial viscosity abnormalities in disease models, highlighting its potential for drug efficacy evaluation. Notably, MPS5 also enables the assessment of acute alcoholic liver injury in mouse models at the organ level.