Neural differentiation on EM grids – iNeurons sample preparation for cryo-ET and CLEM

Output Details

This is a protocol for differentiating AAVS1-TRE3G-NGN2 iPSCs and hESCs to iNeurons directly on EM grids for cryo-ET and cryo-CLEM. The protocol comprises the following steps: (1) EM grids coating with Matrigel or Poly-Ornithine and Laminin; (2) differentiation and seeding of iNeurons on EM grids; (3) lentiviral transduction for transient expression of fluorescently-labelled proteins; (4) plunge freezing.
Tags
  • Cryo-ET
  • Differentiation
  • Human
  • In Cellulo
  • In Vitro
  • iPSC
  • Stem cells – embryonic

Meet the Authors

  • Cristina Capitanio, MSc

    Key Personnel: Team Harper

    Max Planck Institute of Biochemistry

  • Victoria Anne Trinkaus, MSc

    Key Personnel: Team Harper

    Max Planck Institute of Biochemistry

  • Melissa Hoyer, PhD

    Key Personnel: Team Harper

    Harvard University