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Microfluidic Device for High-Resolution Cytoskeleton Imaging and Washout Assays in Physcomitrium (Physcomitrella)
Mari W Yoshida1, Elena Kozgunova2,3
1Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Aichi, Japan.
Methods in Molecular Biology (Clifton, N.J.)
|February 11, 2023
Summary
High-resolution imaging of cytoskeleton dynamics in moss cells using oblique illumination microscopy in microfluidic devices reveals cellular function. Protocols detail sample preparation, microscopy, and chemical treatments for live-cell studies.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Plant Science
Background:
- Cytoskeleton dynamics are crucial for cellular functions and protein interactions.
- Oblique illumination fluorescent microscopy is effective for imaging near the plasma membrane.
- Understanding these dynamics requires high spatiotemporal resolution.
Purpose of the Study:
- To provide detailed protocols for high-resolution cytoskeleton imaging.
- To utilize moss Physcomitrella (Physcomitrium patens) cells within microfluidic devices.
- To enable live-cell imaging with controlled chemical compound introduction and washout.
Main Methods:
- Preparation of polydimethylsiloxane (PDMS) microfluidic devices.
- Culture of moss protonema and gametophore cells.
- Short- and long-term oblique illumination fluorescent microscopy.
- Introduction and washout of chemical compounds (e.g., microtubule-disrupting drugs) during imaging.
Main Results:
- Established protocols for high-resolution imaging of cytoskeleton dynamics in moss cells.
- Demonstrated the utility of microfluidic devices for live-cell imaging of cellular events.
- Enabled dynamic observation of cellular responses to chemical treatments.
Conclusions:
- The developed protocols facilitate detailed studies of cytoskeleton dynamics in Physcomitrella (Physcomitrium patens).
- Microfluidic devices combined with oblique illumination microscopy offer a powerful platform for live-cell imaging.
- This approach provides insights into cellular function and responses to external stimuli.
