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

By Cristina Capitanio, MSc, Victoria Anne Trinkaus, MSc and Melissa Hoyer, PhD
View Published Protocol

Output Details

Description
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.
Identifier (DOI)
10.17504/protocols.io.5jyl8jz36g2w/v2
Tags
  • Cryo-ET
  • Differentiation
  • Human
  • In Cellulo
  • In Vitro
  • iPSC
  • Stem cells – embryonic
Team
Team Harper
Program
Collaborative Research Network
Theme
PD Functional Genomics

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

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