@inbook{9756,
  abstract     = {High-resolution visualization and quantification of membrane proteins contribute to the understanding of their functions and the roles they play in physiological and pathological conditions. Sodium dodecyl sulfate-digested freeze-fracture replica labeling (SDS-FRL) is a powerful electron microscopy method to study quantitatively the two-dimensional distribution of transmembrane proteins and their tightly associated proteins. During treatment with SDS, intracellular organelles and proteins not anchored to the replica are dissolved, whereas integral membrane proteins captured and stabilized by carbon/platinum deposition remain on the replica. Their intra- and extracellular domains become exposed on the surface of the replica, facilitating the accessibility of antibodies and, therefore, providing higher labeling efficiency than those obtained with other immunoelectron microscopy techniques. In this chapter, we describe the protocols of SDS-FRL adapted for mammalian brain samples, and optimization of the SDS treatment to increase the labeling efficiency for quantification of Cav2.1, the alpha subunit of P/Q-type voltage-dependent calcium channels utilizing deep learning algorithms.},
  author       = {Kaufmann, Walter and Kleindienst, David and Harada, Harumi and Shigemoto, Ryuichi},
  booktitle    = { Receptor and Ion Channel Detection in the Brain},
  isbn         = {9781071615218},
  keywords     = {Freeze-fracture replica: Deep learning, Immunogold labeling, Integral membrane protein, Electron microscopy},
  pages        = {267--283},
  publisher    = {Humana},
  title        = {{High-Resolution localization and quantitation of membrane proteins by SDS-digested freeze-fracture replica labeling (SDS-FRL)}},
  doi          = {10.1007/978-1-0716-1522-5_19},
  volume       = {169},
  year         = {2021},
}

@article{8586,
  abstract     = {Cryo-electron microscopy (cryo-EM) of cellular specimens provides insights into biological processes and structures within a native context. However, a major challenge still lies in the efficient and reproducible preparation of adherent cells for subsequent cryo-EM analysis. This is due to the sensitivity of many cellular specimens to the varying seeding and culturing conditions required for EM experiments, the often limited amount of cellular material and also the fragility of EM grids and their substrate. Here, we present low-cost and reusable 3D printed grid holders, designed to improve specimen preparation when culturing challenging cellular samples directly on grids. The described grid holders increase cell culture reproducibility and throughput, and reduce the resources required for cell culturing. We show that grid holders can be integrated into various cryo-EM workflows, including micro-patterning approaches to control cell seeding on grids, and for generating samples for cryo-focused ion beam milling and cryo-electron tomography experiments. Their adaptable design allows for the generation of specialized grid holders customized to a large variety of applications.},
  author       = {Fäßler, Florian and Zens, Bettina and Hauschild, Robert and Schur, Florian KM},
  issn         = {1047-8477},
  journal      = {Journal of Structural Biology},
  keywords     = {electron microscopy, cryo-EM, EM sample preparation, 3D printing, cell culture},
  number       = {3},
  publisher    = {Elsevier},
  title        = {{3D printed cell culture grid holders for improved cellular specimen preparation in cryo-electron microscopy}},
  doi          = {10.1016/j.jsb.2020.107633},
  volume       = {212},
  year         = {2020},
}

@inproceedings{11222,
  author       = {Kim, Olena and Borges Merjane, Carolina and Jonas, Peter M},
  booktitle    = {Intrinsic Activity},
  issn         = {2309-8503},
  keywords     = {hippocampus, mossy fibers, readily releasable pool, electron microscopy},
  location     = {Innsbruck, Austria},
  number       = {Suppl. 1},
  publisher    = {Austrian Pharmacological Society},
  title        = {{Functional analysis of the docked vesicle pool in hippocampal mossy fiber terminals by electron microscopy}},
  doi          = {10.25006/ia.7.s1-a3.27},
  volume       = {7},
  year         = {2019},
}