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Plunge-freezing allows the trapping of potentially complex conformer mixtures for subsequent high-resolution structural analysis by cryogenic electron microscopy. The ultrafast nature of the vitrification process, when combined with a defined activation of the sample, can enable time-resolved structural studies. For light-sensitive samples, this activation can be achieved by controlled illumination of the sample during the plunge-freezing process. Here, we describe a simple approach that enables users to modify commercially available plunge-freezers with a light-emitting diode (LED)-based illumination system, and describe a straightforward protocol for its application to the study of red-light-sensitive phytochromes.

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A Simple Modification of a Commercial Plunge-Freezer to Allow Time-Resolved Vitrification for Cryo-EM Analyses of Light-Activated Proteins

  • 实验方案

  • 发表日期：

    2026年

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  • Andrea Flores-Ibarra,
  • Monika Gunkel,
  • Elmar Behrmann

摘要

Plunge-freezing allows the trapping of potentially complex conformer mixtures for subsequent high-resolution structural analysis by cryogenic electron microscopy. The ultrafast nature of the vitrification process, when combined with a defined activation of the sample, can enable time-resolved structural studies. For light-sensitive samples, this activation can be achieved by controlled illumination of the sample during the plunge-freezing process. Here, we describe a simple approach that enables users to modify commercially available plunge-freezers with a light-emitting diode (LED)-based illumination system, and describe a straightforward protocol for its application to the study of red-light-sensitive phytochromes.