Visualization of cytoskeletal elements in the transmission X-ray microscope

Area of Science:

• Cell Biology
• Microscopy Techniques

Background:

• Cytoskeletal filaments are crucial for cell structure and function.
• Transmission X-ray microscopy (TXM) offers potential for high-resolution cellular imaging.
• Visualizing cytoskeletons in intact cells presents contrast challenges.

Purpose of the Study:

• To investigate the arrangement of cytoskeletal filaments in interphase PtK2 cells using TXM.
• To determine the effectiveness of detergent extraction in improving TXM image contrast for cytoskeletal studies.
• To compare TXM images with transmission electron microscopy (TEM) for cytoskeletal visualization.

Main Methods:

• PtK2 cells were prepared using nonionic detergent Triton X-100 extraction to remove soluble proteins and organelles.
• Transmission X-ray microscopy was employed to image cytoskeletal filaments in both air-dried and hydrated specimens.
• Specimens were also examined using transmission electron microscopy for comparative analysis.

Main Results:

• Detergent extraction was essential for sufficient image contrast, enabling visualization of cytoskeletal filaments.
• TXM images of cytoskeletons and endoplasmic reticulum showed similarities to TEM images.
• TXM achieved a resolution of approximately 50 nm and revealed nuclear substructure in hydrated cells.

Conclusions:

• TXM, particularly after detergent extraction, is a viable technique for studying cytoskeletal arrangements.
• The resolution of TXM allows for the observation of cellular substructures, including nuclear features.
• TXM provides a valuable complementary method to TEM for cell biology research.