Correlative light and electron microscopy at defined cell cycle stages in a controlled environment

Area of Science:

• Cell Biology
• Microscopy
• Biophysics

Background:

• Cells exhibit dynamic architectural changes in response to stimuli.
• Understanding these dynamics at high resolution is crucial for mechanistic insights.
• Current methods may not fully preserve native cellular conditions during observation.

Purpose of the Study:

• To present a novel protocol for in situ study of dynamic intracellular structures.
• To achieve high spatiotemporal resolution under near-native conditions.
• To enable mechanistic understanding of cellular dynamics.

Main Methods:

• Development of a specialized chamber for cell culture, transport, and imaging.
• Environmental control (temperature, gas concentration) within the chamber.
• Application of the protocol to study ultrastructural changes during the mammalian cell cycle.

Main Results:

• Demonstration of a protocol for in situ imaging of dynamic cellular processes.
• Quantification of ultrastructural changes during the cell cycle with high resolution.
• Validation of the environmental control system for maintaining near-native conditions.

Conclusions:

• The developed protocol enables high-resolution, in situ study of cellular dynamics.
• Environmental control is key to preserving native conditions for accurate observation.
• The method is adaptable for primary cells, tissues, and organoids.