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DOI: 10.3791/54874-v
This manuscript describes an easy-to-use and low-cost cryofixation method for visualizing suspension cells by transmission electron microscopy. The technique provides excellent preservation of cellular structure, making it suitable for ultrastructural and immunolocalization studies.
This manuscript describes an easy-to-use and low-cost cryofixation method for visualizing suspension cells by transmission electron microscopy.
The goal of the following experiments is to propose a tool for ultrastructural and immunolocalization studies of suspension cells for visualization in transmission electron microscopy. This technique is easy to use, low cost, and gives excellent preservation of cellular structure. This is a culture of suspension cells.
First, the cells are harvested by different centrifugations to obtain a pellet in a microcentrifuge tube. In a second step, the sample is rapidly frozen by plunge freezing which immobilizes the cells and limits the formation of ice crystals. Next, freeze substitution is performed in order to replace water by solvent containing osmium tetroxide as fixative to stabilize cell structures.
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