Related Experiment Video
Updated: Feb 15, 2026

Micropatterning Transmission Electron Microscopy Grids to Direct Cell Positioning within Whole-Cell Cryo-Electron Tomography Workflows
Published on: September 13, 2021
Electron microscopy of Drosophila garland cell nephrocytes: Optimal preparation, immunostaining and STEM tomography
Florian Hochapfel1,2, Lucia Denk1, Christine Maaßen1
1Molecular and Cellular Anatomy, University of Regensburg, Regensburg, Germany.
Abstract:
Due to its structural and molecular similarities to mammalian podocytes, the Drosophila nephrocyte emerged as a model system to study podocyte development and associated diseases. Similar to podocytes, nephrocytes establish a slit diaphragm between foot process-like structures in order to filter the hemolymph. One major obstacle in nephrocyte research is the distinct visualization of this subcellular structure to assess its integrity. Therefore, we developed a specialized dissection and fixation protocol, including high pressure freezing and freeze substitution techniques, to improve the preservation of the intricate ultrastructural details necessary for electron microscopic assessment. By means of scanning transmission electron microscopy (STEM) tomography, a three-dimensional dataset was generated to further understand the complex architecture of the nephrocyte channel system. Moreover, a staining protocol for immunolabeling of ultrathin sections of Epon-embedded nephrocytes is discussed, which allows the reliable detection of GFP-tagged fusion proteins combined with superior sample preservation. Due to the growing number of available GFP-trap fly lines, this approach is widely applicable for high resolution localization studies in wild type and mutant nephrocytes.
Related Concept Videos
Preparation of Samples for Electron Microscopy
Overview of Electron Microscopy
Scanning Electron Microscopy
Fundamental Principles
Accelerated...
Transmission Electron Microscopy
Immunogold Electron Microscopy
Cryo-electron Microscopy

