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Mitochondrial dysfunction is linked to many neurological diseases; therefore, the ability to measure mitochondrial function is of great use for researching disease and testing potential therapeutics. Here we describe a high-content assay to simultaneously measure mitochondrial membrane potential, morphology, and cell viability in iPSC-derived neurons. Neurons are seeded into plates suitable for fluorescent microscopy, and stained with the mitochondrial membrane potential-dependent dye TMRM, cytoplasmic dye Calcein-AM, and nuclear stain Hoechst-33,342. Images are acquired in live cells and analyzed using automated image analysis software.

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High Content Analysis of Mitochondrial Function in Induced Pluripotent Stem Cell-Derived Neurons

  • 实验方案

  • 发表日期：

    2025年

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  • Daniel Little,
  • Christin Luft,
  • Olukunbi Mosaku,
  • Robin Ketteler,
  • Michael J. Devine,
  • Paul Gissen

摘要

Mitochondrial dysfunction is linked to many neurological diseases; therefore, the ability to measure mitochondrial function is of great use for researching disease and testing potential therapeutics. Here we describe a high-content assay to simultaneously measure mitochondrial membrane potential, morphology, and cell viability in iPSC-derived neurons. Neurons are seeded into plates suitable for fluorescent microscopy, and stained with the mitochondrial membrane potential-dependent dye TMRM, cytoplasmic dye Calcein-AM, and nuclear stain Hoechst-33,342. Images are acquired in live cells and analyzed using automated image analysis software.