A method for observing intracellular structures of free cells by scanning electron microscopy

Area of Science:

• Cell Biology
• Microscopy Techniques
• Materials Science

Background:

• Scanning electron microscopy (SEM) is crucial for visualizing cell ultrastructure.
• Conventional embedding methods for SEM are often incompatible with osmium-based maceration procedures.
• Existing techniques can introduce artifacts or obscure fine intracellular details.

Purpose of the Study:

• To introduce a new embedding method for SEM of free cultured cells.
• To evaluate the efficacy of chitosan embedding combined with the Osmium-DMSO-Osmium (O-D-O) method.
• To demonstrate the preservation of intracellular structures and absence of artifacts.

Main Methods:

• Cultured free cells were embedded using chitosan.
• The Osmium-DMSO-Osmium (O-D-O) procedure was applied to the chitosan-embedded cells.
• Scanning electron microscopy was used for observation of the prepared samples.

Main Results:

• Chitosan embedding is compatible with the O-D-O maceration process.
• Intracellular structures of free cells were clearly visualized.
• Chitosan embedding prevented the destruction of the embedding medium by osmium maceration, unlike other tested media.
• The method produced featureless fracture faces without recognizable artifacts.

Conclusions:

• Chitosan embedding offers a robust and artifact-free method for SEM of cultured free cells.
• This technique enhances the observation of intracellular details previously obscured by embedding media degradation.
• The Osmium-DMSO-Osmium (O-D-O) method combined with chitosan embedding is a valuable advancement in cell microscopy.