Electron Microscopy of Cells Grown on Polyacrylamide Hydrogels
Frederik Helmprobst1,2, Anna Hupfer3, Matthias Lauth3
1Philipps University Marburg, Institute of Neuropathology, Marburg, Germany.
Current Protocols
|August 9, 2022
Summary
Researchers developed a new method to embed cells grown on stiff polyacrylamide hydrogels for electron microscopy. This technique simplifies processing, enabling better visualization of cells in a more native environment.
Area of Science:
- Cell Biology
- Materials Science
- Microscopy
Background:
- Standard cell culture substrates like plastic and glass are significantly stiffer than physiological tissues, leading to artificial cell behavior.
- Physiologically relevant stiffness (<100 kPa) can be achieved using polymer-based hydrogels for 2D and 3D cell culture.
- Polyacrylamide hydrogels offer tunable properties and are compatible with various downstream biological assays.
Purpose of the Study:
- To address challenges in processing high-stiffness polyacrylamide hydrogels for electron microscopy.
- To develop a simplified protocol for embedding cells cultured on these hydrogels without altering standard procedures.
Main Methods:
- Development of a novel protocol for embedding cells grown on high-stiffness polyacrylamide hydrogels.
- Adaptation of routine embedding protocols for transmission electron microscopy.
Main Results:
- Successful embedding and processing of cells cultured on high-stiffness polyacrylamide hydrogels.
- The new protocol does not necessitate modifications to standard electron microscopy embedding techniques.
Conclusions:
- The developed protocol facilitates the electron microscopy analysis of cells cultured on stiff polyacrylamide hydrogels.
- This method allows for the study of cells in a more native in vitro environment using electron microscopy.
